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WO2024249967A1 - Rna vaccines encoding herpes simplex virus glycoproteins and uses thereof - Google Patents

Rna vaccines encoding herpes simplex virus glycoproteins and uses thereof Download PDF

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Publication number
WO2024249967A1
WO2024249967A1 PCT/US2024/032163 US2024032163W WO2024249967A1 WO 2024249967 A1 WO2024249967 A1 WO 2024249967A1 US 2024032163 W US2024032163 W US 2024032163W WO 2024249967 A1 WO2024249967 A1 WO 2024249967A1
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Prior art keywords
hsv
seq
immunogenic fragment
sequence
amino acid
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French (fr)
Inventor
Harvey Friedman
Gary H. Cohen
Sita Awasthi
Drew Weissman
Kevin EGAN
John Lubinski
Alptekin GÜLER
Ricardo Sanchez Velazquez
Julia UEBELE
Annette VOGEL
Christina PFAFENROT
Anna Luise Ernst
Stephanie HEIN
Sabrina Hinz
Sarah Catharina Dany
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University of Pennsylvania Penn
Biontech US Inc
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University of Pennsylvania Penn
Biontech US Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A61P31/22—Antivirals for DNA viruses for herpes viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/53—DNA (RNA) vaccination
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/16011—Herpesviridae
    • C12N2710/16611—Simplexvirus, e.g. human herpesvirus 1, 2
    • C12N2710/16622—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/16011—Herpesviridae
    • C12N2710/16611—Simplexvirus, e.g. human herpesvirus 1, 2
    • C12N2710/16634—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • compositions for the prevention and treatment of genital herpes comprising nucleoside modified RNAs that encode herpes simplex virus (HSV) glycoproteins, including those involved in virus entry and immune evasion, and methods of use thereof.
  • HSV herpes simplex virus
  • BACKGROUND Herpes simplex type 1 (HSV-1) infections affect an estimated 3.7 billion people between the ages of 0-49 worldwide. Many people acquire HSV-1 during childhood from family members, friends, or peers at school. Orolabial HSV-1 infection is usually self-limited and although emotionally disturbing to some, does not pose a serious health risk to those infected.
  • HSV-1 infection at other sites may cause blindness and life- threatening encephalitis, respectively.
  • reduced orolabial HSV-1 acquisition during childhood has resulted in adolescents and adults having a higher risk of acquiring first-time HSV-1 as a genital infection.
  • genital HSV- 1 infections have a reduced risk of recurrences, genital transmission of HSV-1 from mother to newborn during labor 1 11979815v1 P-627574-PC and delivery can be devastating.
  • HSV-1 neonatal herpes is more common than HSV-2 neonatal herpes in resource rich-countries.
  • compositions e.g., immunogenic compositions, e.g., vaccines
  • HSV herpes simplex virus
  • immunogenic fragment thereof e.g., HSV-1 glycoprotein or immunogenic fragment thereof, an HSV-2 glycoprotein or immunogenic fragment thereof, or any combinations thereof
  • subject e.g., a patient
  • related technologies e.g., methods
  • the present disclosure provides HSV (e.g., HSV-1, HSV-2, or both) vaccine compositions and related technologies (e.g., methods). Additionally, the present disclosure provides HSV (e.g., HSV-1, HSV-2, or both) vaccine compositions for inhibiting, preventing or treating an HSV infection (e.g., HSV-1, HSV-2, or both).
  • HSV e.g., HSV-1, HSV-2, or both
  • vaccine compositions for inhibiting, preventing or treating an HSV infection (e.g., HSV-1, HSV-2, or both).
  • the present disclosure provides a method of inhibiting an HSV-1 oral mucosal infection in a subject comprising the step of administering to said subject a composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, a composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and a composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof.
  • HSV Herpes Simplex Virus
  • gC Herpes Simplex Virus
  • gE HSV glycoprotein E
  • the present disclosure provides a method of preventing establishment of a latent HSV infection in a subject comprising the step of administering to said subject a composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, a composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and a composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof.
  • HSV Herpes Simplex Virus
  • gC Herpes Simplex Virus
  • gE HSV glycoprotein E
  • Figures 1A-1C Characterization of exemplary translational products of an immunogenic fragment of the ectodomain from HSV-2 gC (gC2)-, HSV-2 gD (gD2)- and HSV-2 gE (gE2)- modified RNA in Vero cells.
  • Figure 1A Western blot showing expression of an immunogenic fragment of gC2 from an exemplary modified RNA.
  • Figure 1B Western blot showing expression of an immunogenic fragment of gD2 from an exemplary modified RNA.
  • Figure 1C Western blot showing expression of an immunogenic fragment of gE2 from an exemplary modified RNA.
  • Figure 2A Western blot showing expression of an immunogenic fragment of gE2 from an exemplary modified RNA.
  • gC2 antibody (Ab) response as determined by antigen-specific ELISA in mice immunized with an exemplary gD2 RNA; or an exemplary combination of a gC2 RNA, a gD2 RNA and a gE2 RNA each given at a different intradermal site (Trivalent-I); or an exemplary combination of a gC2 RNA, a gD2 RNA and a gE2 RNA given in a single composition (Trivalent- C). I indicates first immunization; II indicates second immunization. [0016] Figure 2B.
  • gE2 Ab response as determined by antigen-specific ELISA in mice immunized with an exemplary gD2 RNA; or an exemplary combination of a gC2 RNA, a gD2 RNA and a gE2 RNA each given at a different intradermal site (Trivalent-I); or an exemplary 3 11979815v1 P-627574-PC combination of a gC2 RNA, gD2 RNA and gE2 RNA given in a single composition (Trivalent- C). I indicates first immunization; II indicates second immunization. [0018] Figure 3A. Antigen-specific IgG1 responses in mice vaccinated with exemplary RNA compositions.
  • Antibodies were evaluated after the first and second immunization for IgG1 responses. I indicates first immunization; II indicates second immunization.
  • Figure 3B Antigen-specific IgG2a responses in mice vaccinated with exemplary RNA compositions. Antibodies were evaluated after the first and second immunization for IgG2a responses. I indicates first immunization; II indicates second immunization.
  • Figure 4. Neutralizing antibody titers in RNA vaccinated mice. 50% endpoint neutralization titers of serum were obtained after the second immunization. Titers were performed using 10% human complement.
  • Trivalent-I animals were immunized with an exemplary gC2/lipid nanoparticle (LNP), an exemplary gD2/LNP, and an exemplary gE2/LNP, each given at a different site.
  • Trivalent-C animals were immunized with an exemplary gC2, an exemplary gD2 and an exemplary gE2 combined into a single LNP.
  • Figures 5A-5B Figures 5A-5B.
  • Splenocytes were stimulated with subunit antigen glycoproteins (Figure 5A) or 15 amino acid peptides with 11 overlapping amino acids to stimulate HSV-2 specific T cell responses (Figure 5B). * indicates p ⁇ 0.05 (t test) comparing gC, gD or gE with PBS stimulated CD4 + T cells or DMSO stimulated CD4 + T cells. Error bars represent SD. [0022] Figures 6A-6B.
  • Trivalent-I represents animals immunized with an exemplary gC2 RNA/LNP, an exemplary gD2 RNA /LNP, and an exemplary gE2 RNA /LNP each given at different intradermal sites.
  • Trivalent-C represents 4 11979815v1 P-627574-PC animals immunized with an exemplary gC2 RNA, an exemplary gD2 RNA and an exemplary gE2 RNA combined into a single LNP for immunization.
  • Figure 7B Weight loss (-) or gain (+) and neurological signs in BALB/c mice immunized with exemplary RNA compositions twice at 28 day intervals and challenged intravaginally with HSV-2.
  • Trivalent-I represents animals immunized with an exemplary gC2 RNA/LNP, an exemplary gD2 RNA /LNP, and an exemplary gE2 RNA /LNP each given at different intradermal sites.
  • Trivalent-C represents animals immunized with an exemplary gC2 RNA, an exemplary gD2 RNA and an exemplary gE2 RNA combined into a single LNP for immunization.
  • Figures 8A-8B Vaginal viral titers in mice vaccinated with exemplary RNA compositions after intravaginal challenge with HSV-2.
  • FIG. 8A Vaginal swab titers were obtained on Day 2 ( Figure 8A) and Day 4 (Figure 8B) post-challenge. The dotted lines indicate the limit of detection of the assay at 7 PFU/ml.
  • Figure 9 Genital disease in mice vaccinated with exemplary RNA compositions after HSV-2 vaginal challenge. Mice were immunized with poly C as a control, or with an exemplary gD2 RNA/LNP, trivalent at individual sites for each exemplary glycoprotein RNA (Trivalent-I) or trivalent with all three exemplary RNAs combined (Trivalent-C). Genital disease was scored for 28 days. All animals in the poly C group died by day 10.
  • FIG. 10 HSV-2 DNA copies in dorsal root ganglia (DRG) in RNA vaccinated mice on day 4 post challenge. HSV-2 DNA in DRG was measured by qPCR. DRG from 4 to 5 animals per group were evaluated for HSV-2 DNA at 4 days post challenge. The bars represent the mean values per group.
  • Figure 11A The exemplary trivalent RNA-LNP vaccine induces a potent T follicular helper cell response in mice.
  • BALB/c female mice were left un-immunized as na ⁇ ve control animals or immunized intradermally twice at 28 day intervals with poly C RNA-LNP or an exemplary trivalent modified RNA-LNP.
  • the poly C RNA controls received 10 ⁇ g Poly C RNA- LNP divided into 4 aliquots and administered at 4 separate sites.
  • the exemplary trivalent modified RNA group received 10 ⁇ g of an exemplary gC2 RNA-LNP, 10 ⁇ g of an exemplary gD2 RNA- LNP, and 10 ⁇ g of an exemplary gE2 RNA-LNP each divided into 2 aliquots and each given at 2 sites.
  • RNA-LNP vaccine induces a potent germinal center B cell response in mice.
  • BALB/c female mice were left un-immunized as na ⁇ ve control animals or immunized intradermally twice at 28 day intervals with poly C RNA-LNP or an exemplary trivalent modified RNA-LNP.
  • the poly C RNA controls received 10 ⁇ g Poly C RNA- LNP divided into 4 aliquots and administered at 4 separate sites.
  • the trivalent modified RNA group received 10 ⁇ g of an exemplary gC2 RNA-LNP, 10 ⁇ g of an exemplary gD2 RNA-LNP, and 10 ⁇ g of an exemplary gE2 RNA-LNP each divided into 2 aliquots and each given at 2 sites. Two weeks after the second immunization, spleens were harvested from 5 animals per group and flow cytometry performed to detect germinal center B cell responses (*p ⁇ 0.05). [0030] Figures 12A-12C. Genital mucosa IgG antibody responses.
  • mice were immunized intradermally twice at 28 day intervals with 10 ⁇ g of poly C RNA-LNP, 10 ⁇ g of an exemplary gD2 RNA-LNP or 10 ⁇ g each of an exemplary gC2, an exemplary gD2, an exemplary gE2 trivalent modified RNA-LNP.
  • the exemplary trivalent RNA was combined and administered as 10 ⁇ g of an exemplary gC2 RNA & 10 ⁇ g of an exemplary gD2 RNA & 10 ⁇ g of an exemplary gE2 RNA combined into LNP and divided into 4 aliquots and given at 4 sites.
  • 60 ⁇ l of media was introduced in the vaginal cavity and retrieved.
  • Figure 13 The exemplary trivalent RNA-LNP vaccine produces antibodies that block gC2 binding to complement component C3b.
  • mice were left unimmunized as a source of non-immune IgG, or immunized intradermally with poly C RNA-LNP or an exemplary trivalent RNA-LNP.
  • the poly C RNA controls received 10 ⁇ g poly C RNA-LNP divided into 4 aliquots and administered at 4 separate sites.
  • the gD2 RNA group received 10 ⁇ g of an exemplary gD2 RNA- LNP administered as described for the poly C RNA-LNP.
  • the trivalent modified RNA group received 10 ⁇ g of an exemplary gC2 RNA-LNP, 10 ⁇ g of an exemplary gD2 RNA-LNP, and 10 ⁇ g of an exemplary gE2 RNA-LNP combined into LNP and divided into 4 aliquots and given at 4 sites. There were 10 mice in each group. Sera from the 10 mice were pooled and IgG was purified. The IgG was evaluated at 12 ⁇ g/200 ⁇ l for its ability to block complement component C3b binding to gC2. (****p ⁇ 0.0001). [0032] Figures 14A-14F.
  • the exemplary trivalent RNA vaccine provides outstanding protection in mice when the vaccine is administered intramuscularly.
  • mice were 6 11979815v1 P-627574-PC immunized intramuscularly with poly C RNA-LNP as a control (15/group) or with an exemplary trivalent RNA composition containing 10 ⁇ g each of an exemplary gC2, an exemplary gD2 and an exemplary gE2 RNA-LNP (20/group).
  • Figure 14A presents data on mouse survival;
  • Figure 14B presents data on weight loss;
  • Figure 14C presents data on genital disease.
  • DRG were harvested from nine poly C animals at the time of euthanasia between days 7 and 12 post-infection or at the end of the experiment on day 28 in the trivalent RNA group.
  • Figure 14D presents data on HSV-2 DNA in DRG.
  • Figure 14E presents data on vaginal viral cultures on Day 2 and Figure 14F presents data on vaginal viral cultures on Day 4. Difference between poly C and trivalent groups are significant, p ⁇ 0.001 for Figures 14A-14F.
  • FIG. 15A presents data on survival;
  • Figure 15B provides data on vaginal disease (acute phase); and
  • Figure 15C provides data on vaginal disease (recurrent phase).
  • Exemplary tri-HSV-1 and exemplary tri-HSV-2 immunizations generate antibodies that bind to HSV-1 proteins and neutralize HSV-1.
  • FIG. 16A-16C IgG ELISA endpoint titers against HSV-1 proteins.
  • Figure 16D HSV-1 neutralizing titers in the presence of 5% human serum as source of complement.
  • N 10 mice per group. P values were calculated using the two-tailed Mann-Whitney test.
  • FIG. 18A Single cytokine positive CD8+ T-cells were gated using CD8+ and TNF ⁇ , IFN ⁇ , or IL-2 parameters. Gate coordinates were set based on the DMSO treated cells and those gate coordinates were transferred to the peptide treated cells.
  • Figures 18A-18D Exemplary tri-HSV-1 and exemplary tri-HSV-2 immunizations generate CD4+ and CD8+ T-cell responses to HSV-1 overlapping peptides. Mice were 7 11979815v1 P-627574-PC immunized twice with 10 ⁇ g total of exemplary tri-HSV-1 RNA, exemplary tri-HSV-2 RNA, or PBS.
  • Figure 18A Exemplary tri-HSV-1 and exemplary tri-HSV-2 immunizations generate CD4+ and CD8+ T-cell responses to HSV-1 overlapping peptides. Mice were 7 11979815v1 P-627574-PC immunized twice with 10 ⁇ g total of exemplary tri-HSV-1
  • Figure 18B Activation markers of CD8+ T-cells stimulated with gE1 overlapping peptide pool producing TNF ⁇ , IFN ⁇ , and/or IL-2.
  • Figure 18C Proportion of CD8+ T-cells expressing CD107a when stimulated with gE1 overlapping peptides.
  • FIGS 19A-19G Exemplary tri-HSV-1 and tri-HSV-2 RNA compositions protect mice against acute and latent HSV-1 infection in the lip. Mice were immunized with PBS, exemplary tri-HSV-1 RNA, or tri-HSV-2 RNA compositions as in Figure 1. Mice were challenged with 2 ⁇ 10 6 PFU of HSV-1 applied to the scarified lower lip.
  • Figure 19A HSV-1 virus titers in the lower lip.
  • Figure 19B HSV-1 DNA copy number in the lower lip.
  • Figure 19C Figure 19C.
  • FIG. 19D HSV-1 virus titers in the trigeminal ganglia
  • Figure 19D HSV-1 DNA copy number in the TG.
  • Figures 19A-19D. n 10 per group.
  • Figure 19E. Mice were weighed daily for 13 days.
  • Figure 19F. Mice were scored for lesions on the lip with a score of 1 for lesions or 0 for no lesions.
  • Figure 19G. HSV-1 DNA genome copies in the TG 28 days post-challenge.
  • P values in Figures 19A-19D, and Figure 19G were calculated by the two-tailed Mann-Whitney test. [0038] Figures 20A-20F.
  • Exemplary tri-HSV-1 and exemplary tri-HSV-2 RNA compositions protect mice from intravaginal HSV-1 disease.
  • Mice were immunized with PBS, exemplary tri- HSV-1 RNA, or exemplary tri-HSV-2 RNA.
  • Mice were challenged with 2 ⁇ 10 6 PFU of HSV-1 intravaginally and scored for clinical disease for 14 days.
  • Figure 20A Survival curve. P values were calculated by the log-rank test.
  • Figure 20C Mean genital disease scores for each group.
  • Figures 20D-20E Vaginal swabs were obtained on days two and four for virus titers.
  • Figure 20F HSV-1 DNA copy number in DRG.
  • Trivalent HSV-2 vaccine (Tri-HSV-2) protects BALB/c mice against HSV-1 encephalitis.
  • Female BALB/c mice were immunized twice with either 1 ⁇ g (0.33 ⁇ g of each mRNA) or 10 ⁇ g (3.33 ⁇ g each mRNA) trivalent HSV-2 mRNA vaccine at 1-month intervals or PBS as described above.
  • agent may refer to a physical entity or phenomenon. In some embodiments, an agent may be characterized by a particular feature and/or effect. In some 9 11979815v1 P-627574-PC embodiments, an agent may be a compound, molecule, or entity of any chemical class including, for example, a small molecule, polypeptide, nucleic acid, saccharide, lipid, metal, or a combination or complex thereof. In some embodiments, the term “agent” may refer to a compound, molecule, or entity that comprises a polymer. In some embodiments, the term may refer to a compound or entity that comprises one or more polymeric moieties.
  • the term “agent” may refer to a compound, molecule, or entity that is substantially free of a particular polymer or polymeric moiety. In some embodiments, the term may refer to a compound, molecule, or entity that lacks or is substantially free of any polymer or polymeric moiety.
  • Amino acid In its broadest sense, as used herein, the term “amino acid” refers to a compound and/or substance that can be, is, or has been incorporated into a polypeptide chain, e.g., through formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H 2 N–C(H)(R)–COOH.
  • an amino acid is a naturally- occurring amino acid. In some embodiments, an amino acid is a non-natural amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L- amino acid. “Standard amino acid” refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. “Nonstandard amino acid” refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. In some embodiments, an amino acid, including a carboxy- and/or amino-terminal amino acid in a polypeptide, can contain a structural modification as compared with the general structure above.
  • an amino acid may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and/or substitution (e.g., of the amino group, the carboxylic acid group, one or more protons, and/or the hydroxyl group) as compared with the general structure.
  • such modification may, for example, alter the circulating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino acid.
  • such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino acid, as compared with one containing an otherwise identical unmodified amino acid.
  • the term “amino acid” may be used to refer to a free amino acid; in some embodiments it may be used to refer to an amino acid residue of a polypeptide.
  • Antibody agent refers to an agent that specifically binds to a particular antigen. In some embodiments, the term encompasses a 10 11979815v1 P-627574-PC polypeptide or polypeptide complex that includes immunoglobulin structural elements sufficient to confer specific binding.
  • an antibody agent is or comprises a polypeptide whose amino acid sequence includes one or more structural elements recognized by those skilled in the art as a complementarity determining region (CDR); in some embodiments an antibody agent is or comprises a polypeptide whose amino acid sequence includes at least one CDR (e.g., at least one heavy chain CDR and/or at least one light chain CDR) that is substantially identical to one found in a reference antibody. In some embodiments an included CDR is substantially identical to a reference CDR in that it is either identical in sequence or contains between 1-5 amino acid substitutions as compared with the reference CDR.
  • CDR complementarity determining region
  • an included CDR is substantially identical to a reference CDR in that it shows at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that it shows at least 96%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that at least one amino acid within the included CDR is deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR.
  • an included CDR is substantially identical to a reference CDR in that 1-5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that at least one amino acid within the included CDR is substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that 1-5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR.
  • an antibody agent is or comprises a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art as an immunoglobulin variable domain.
  • an antibody agent is a polypeptide protein having a binding domain which is homologous or largely homologous to an immunoglobulin- binding domain.
  • an antibody agent may be or comprise a polyclonal antibody preparation.
  • an antibody agent may be or comprise a monoclonal antibody preparation.
  • an antibody agent may include one or more constant region sequences that are characteristic of a particular organism, such as a camel, human, mouse, primate, rabbit, rat; in many embodiments, an antibody agent may include one or more constant region sequences that are characteristic of a human.
  • an antibody agent may include one or more sequence elements that would be recognized by one skilled in the art as a humanized sequence, a primatized sequence, a chimeric sequence, etc.
  • an antibody agent may be a canonical antibody (e.g., may comprise two heavy chains and two light chains).
  • an antibody agent may be in a format selected from, but not limited to, intact IgA, IgG, IgE or IgM antibodies; bi- or multi- specific antibodies (e.g., Zybodies®, etc); antibody fragments such as Fab fragments, Fab’ fragments, F(ab’)2 fragments, Fd’ fragments, Fd fragments, and isolated CDRs or sets thereof; single chain Fvs; polypeptide-Fc fusions; single domain antibodies (e.g., shark single domain antibodies such as IgNAR or fragments thereof); cameloid antibodies; masked antibodies (e.g., Probodies®); Small Modular ImmunoPharmaceuticals (“SMIPs TM” ); single chain or Tandem diabodies (TandAb®); VHHs; Anticalins®; Nanobodies® minibodies; BiTE®s; ankyrin repeat proteins or DARPINs®; Avimers®; DARTs; TCR-like antibodies;, Adnectins
  • an antibody may lack a covalent modification (e.g., attachment of a glycan) that it would have if produced naturally.
  • an antibody may contain a covalent modification (e.g., attachment of a glycan, a payload (e.g., a detectable moiety, a therapeutic moiety, a catalytic moiety, etc.), or other pendant group (e.g., poly-ethylene glycol, etc.)).
  • Antigen refers to a molecule that is recognized by the immune system, e.g., in particular embodiments the adaptive immune system, such that it elicits an antigen-specific immune response.
  • an antigen-specific immune response may be or comprise generation of antibodies and/or antigen-specific T cells.
  • an antigen is a 12 11979815v1 P-627574-PC peptide or polypeptide that comprises at least one epitope against which an immune response can be generated.
  • an antigen is presented by cells of the immune system such as antigen presenting cells like dendritic cells or macrophages.
  • an antigen or a processed product thereof such as a T-cell epitope is bound by a T- or B-cell receptor, or by an immunoglobulin molecule such as an antibody. Accordingly, an antigen or a processed product thereof may react specifically with antibodies or T lymphocytes (T cells).
  • an antigen is a parasitic antigen.
  • an antigen may be delivered by RNA molecules as described herein.
  • a peptide or polypeptide antigen can be 2-100 amino acids, including for example, 5 amino acids, 10 amino acids, 15 amino acids, 20 amino acids, 25 amino acids, 30 amino acids, 35 amino acids, 40 amino acids, 45 amino acids, or 50 amino acids in length.
  • a peptide or polypeptide antigen can be greater than 50 amino acids. In some embodiments, a peptide or polypeptide antigen can be greater than 100 amino acids.
  • an antigen is recognized by an immune effector cell. In some embodiments, an antigen if recognized by an immune effector cell is able to induce in the presence of appropriate co-stimulatory signals, stimulation, priming and/or expansion of the immune effector cell carrying an antigen receptor recognizing the antigen. In the context of the embodiments of the present disclosure, in some embodiments, an antigen can be presented or present on the surface of a cell, e.g., an antigen presenting cell.
  • an antigen is presented by a diseased cell such as a virus- infected cell.
  • an antigen receptor is a TCR which binds to an epitope of an antigen presented in the context of MHC.
  • binding of a TCR when expressed by T cells and/or present on T cells to an antigen presented by cells such as antigen presenting cells results in stimulation, priming and/or expansion of said T cells.
  • binding of a TCR when expressed by T cells and/or present on T cells to an antigen presented on diseased cells results in cytolysis and/or apoptosis of the diseased cells, wherein said T cells preferably release cytotoxic factors, e.g., perforins and granzymes.
  • cytotoxic factors e.g., perforins and granzymes.
  • a particular entity e.g., polypeptide, genetic signature, metabolite, microbe, etc
  • a particular disease, disorder, or condition if its presence, level and/or form correlates with incidence of, susceptibility to, severity of, stage of, etc. the disease, disorder, or condition (e.g., across a relevant population).
  • two or more 13 11979815v1 P-627574-PC entities are physically “associated” with one another if they interact, directly or indirectly, so that they are and/or remain in physical proximity with one another.
  • two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non-covalently associated, for example by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof.
  • binding typically refers to a non-covalent association between or among entities or moieties. In some embodiments, binding data are expressed in terms of “IC50”.
  • IC50 is the concentration of an assessed agent in a binding assay at which 50% inhibition of binding of reference agent known to bind the relevant binding partner is observed.
  • assays are run under conditions in which the assays are run (e.g., limiting binding target and reference concentrations), these values approximate K D values.
  • Assays for determining binding are well known in the art and are described in detail, for example, in PCT publications WO 94/20127 and WO 94/03205, and other publications such Sidney et al., Current Protocols in Immunology 18.3.1 (1998); Sidney, et al., J. Immunol. 154:247 (1995); and Sette, et al., Mol.
  • binding can be expressed relative to binding by a reference standard peptide.
  • a reference standard peptide For example, can be based on its IC50, relative to the IC50 of a reference standard peptide.
  • Binding can also be determined using other assay systems including those using: live cells (e.g., Ceppellini et al., Nature 339:392 (1989); Christnick et al., Nature 352:67 (1991); Busch et al., Int. Immunol. 2:443 (1990); Hill et al., J. Immunol. 147:189 (1991); del Guercio et al., J. Immunol.
  • Cap refers to a structure comprising or essentially consisting of a nucleoside-5 '-triphosphate that is typically joined to a 5'-end of an uncapped RNA 14 11979815v1 P-627574-PC (e.g., an uncapped RNA having a 5'- diphosphate).
  • a cap is or comprises a guanine nucleotide.
  • a cap is or comprises a naturally-occurring RNA 5’ cap, including, e.g., but not limited to a 7- methylguanosine cap, which has a structure designated as “m7G.”
  • a cap is or comprises a synthetic cap analog that resembles an RNA cap structure and possesses the ability to stabilize RNA if attached thereto, including, e.g., but not limited to anti-reverse cap analogs (ARCAs) known in the art).
  • ARCAs anti-reverse cap analogs
  • a capped RNA may be obtained by in vitro capping of RNA that has a 5' triphosphate group or RNA that has a 5' diphosphate group with a capping enzyme system (including, e.g., but not limited to vaccinia capping enzyme system or Saccharomyces cerevisiae capping enzyme system).
  • a capped RNA can be obtained by in vitro transcription (IVT) of a single-stranded DNA template in the presence of a dinucleotide or trinucleotide cap analog.
  • Cell-mediated immunity “Cell-mediated immunity,” “cellular immunity,” “cellular immune response,” or similar terms are meant to include a cellular response directed to cells characterized by expression of an antigen, in particular characterized by presentation of an antigen with class I or class II MHC.
  • a cellular response relates to immune effector cells, in particular to T cells or T lymphocytes which act as either “helpers” or “killers.”
  • the helper T cells also termed CD4 + T cells or CD4 T cells
  • the killer cells also termed cytotoxic T cells, cytolytic T cells, CD8 + T cells, CD8 T cells, or CTLs kill diseased cells such as virus-infected cells, preventing the production of more diseased cells.
  • co-administration refers to use of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) described herein and an additional therapeutic agent.
  • a pharmaceutical composition e.g., immunogenic composition, e.g., vaccine
  • an additional therapeutic agent may be performed concurrently or separately (e.g., sequentially in any order).
  • a pharmaceutical composition e.g., immunogenic composition, e.g., vaccine
  • an additional therapeutic agent may be combined in one pharmaceutically-acceptable carrier, or they may be placed in separate carriers and delivered to a target cell or administered to a subject at different times.
  • Codon-optimized refers to alteration of codons in a coding region of a nucleic acid molecule to reflect the typical codon usage of a host organism without preferably altering the amino acid sequence encoded by the nucleic acid molecule.
  • coding regions are codon-optimized for optimal expression in a subject to be treated using the RNA molecules described herein.
  • codon-optimization may be performed such that codons for which frequently occurring tRNAs are available are inserted in place of “rare codons.”
  • codon-optimization may include increasing guanosine/cytosine (G/C) content of a coding region of RNA described herein as compared to the G/C content of the corresponding coding sequence of a wild type RNA, wherein the amino acid sequence encoded by the RNA is preferably not modified compared to the amino acid sequence.
  • G/C guanosine/cytosine
  • Combination therapy refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents).
  • the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all “doses” of a first regimen are administered prior to administration of any doses of a second regimen); in some embodiments, such agents are administered in overlapping dosing regimens.
  • “administration” of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modality(ies) in the combination.
  • Comparable refers to two or more agents, entities, situations, sets of conditions, etc., that may not be identical to one another but that are sufficiently similar to permit comparison there between so that one skilled in the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed.
  • comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features.
  • corresponding to may be used to designate the position/identity of a structural element in a compound or composition relative to another compound or composition (e.g., to an appropriate reference compound or composition).
  • a monomeric residue in a polymer e.g., an amino acid residue in a polypeptide or a nucleic acid residue in a polynucleotide
  • corresponding to a residue in an appropriate reference polymer.
  • residues in a polypeptide are often designated using a canonical numbering system based on a reference related polypeptide, so that an amino acid “corresponding to” a residue at position 190, for example, need not actually be the 190 th amino acid in a particular amino acid chain but rather corresponds to the residue found at 190 in the reference polypeptide; those of ordinary skill in the art readily appreciate how to identify “corresponding” amino acids.
  • sequence alignment strategies including software programs such as, for example, BLAST, CS- BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH/GLSEARCH, Genoogle, HMMER, HHpred/HHsearch, IDF, Infernal, KLAST, USEARCH, parasail, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE that can be utilized, for example, to identify “corresponding” residues in polypeptides and/or nucleic acids in accordance with the present disclosure.
  • software programs such as, for example, BLAST, CS- BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH/GLSEARCH, Genoogle, HMMER, HHpred/HHsearch, IDF, Infernal, KLAST, USEARCH, parasail, PSI-BLAST, PSI-Search,
  • corresponding to may be used to describe an event or entity that shares a relevant similarity with another event or entity (e.g., an appropriate reference event or entity).
  • a gene or protein in one organism may be described as “corresponding to” a gene or protein from another organism in order to indicate, in some embodiments, that it plays an analogous role or performs an analogous function and/or that it shows a particular degree of sequence identity or homology, or shares a particular characteristic sequence element.
  • amino acid sequence derived from a designated amino acid sequence (peptide or polypeptide) “derived from” a designated amino acid sequence (peptide or polypeptide), it refers to a structural analogue of a designated amino acid sequence.
  • an amino acid sequence which is derived from a particular amino acid sequence has an amino acid sequence that is identical, essentially identical or homologous to that particular sequence or a fragment thereof.
  • Amino acid sequences derived from a particular amino acid sequence may be variants of that particular sequence or a fragment thereof.
  • the antigens suitable for use herein may be altered such that they vary in sequence from the naturally occurring or native sequences from which they were derived, while retaining the desirable activity of the native sequences.
  • the term “designed” refers to an agent (i) whose structure is or was selected by the hand of man; (ii) that is produced by a process requiring the hand of man; and/or (iii) that is distinct from natural substances and other known agents.
  • Dosing regimen may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time.
  • a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses.
  • a dosing regimen comprises a plurality of doses each of which is separated in time from other doses.
  • individual doses are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses.
  • all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen).
  • Engineered refers to the aspect of having been manipulated by the hand of man.
  • a polynucleotide is considered to be “engineered” when two or more sequences that are not linked together in that order in nature are manipulated 18 11979815v1 P-627574-PC by the hand of man to be directly linked to one another in the engineered polynucleotide and/or when a particular residue in a polynucleotide is non-naturally occurring and/or is caused through action of the hand of man to be linked with an entity or moiety with which it is not linked in nature.
  • Epitope refers to a moiety that is specifically recognized by an immunoglobulin (e.g., antibody or receptor) binding component.
  • an epitope may be recognized by a T cell, a B cell, or an antibody.
  • an epitope is comprised of a plurality of chemical atoms or groups on an antigen.
  • such chemical atoms or groups are surface-exposed when the antigen adopts a relevant three-dimensional conformation.
  • such chemical atoms or groups are physically near to each other in space when the antigen adopts such a conformation.
  • an epitope of an antigen may include a continuous or discontinuous portion of the antigen.
  • an epitope is or comprises a T cell epitope.
  • an epitope may have a length of about 5 to about 30 amino acids, or about 10 to about 25 amino acids, or about 5 to about 15 amino acids, or about 5 to 12 amino acids, or about 6 to about 9 amino acids.
  • a gene product can be a transcript.
  • a gene product can be a polypeptide.
  • expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, etc); (3) translation of an RNA into a polypeptide or protein; and/or (4) post-translational modification of a polypeptide or protein.
  • Five prime untranslated region refers to a sequence of an RNA molecule between a transcription start site and a start codon of a coding region of an RNA.
  • “5’ UTR” refers to a sequence of an RNA molecule that begins at a transcription start site and ends one nucleotide (nt) before a start codon (usually AUG) of a coding region of an RNA molecule, e.g., in its natural context.
  • Humoral immunity As used herein, the term “humoral immunity” or “humoral immune response” refers to antibody production and the accessory processes that accompany it, including: Th2 activation and cytokine production, germinal center formation and isotype switching, affinity 19 11979815v1 P-627574-PC maturation and memory cell generation. It also refers to the effector functions of antibodies, which include pathogen neutralization, classical complement activation, and opsonin promotion of phagocytosis and pathogen elimination.
  • Identity As used herein, the term “identity” refers to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules.
  • polynucleotide molecules e.g., DNA molecules and/or RNA molecules
  • polypeptide molecules are considered to be “substantially identical” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical.
  • Calculation of the percent identity of two nucleic acid or polypeptide sequences for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequence for optimal alignment and non-identical sequences can be disregarded for comparison purposes).
  • the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of a reference sequence.
  • the nucleotides at corresponding positions are then compared. When a position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, then the molecules are identical at that position.
  • the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences.
  • the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm.
  • the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which has been incorporated into the ALIGN program (version 2.0).
  • nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
  • Immunologically equivalent means that an immunologically equivalent molecule such as the immunologically equivalent amino acid sequence exhibits the same or essentially the same immunological properties and/or exerts the 20 11979815v1 P-627574-PC same or essentially the same immunological effects, e.g., with respect to the type of the immunological effect.
  • immunologically equivalent is used with respect to the immunological effects or properties of antigens or antigen variants used for immunization.
  • an amino acid sequence is immunologically equivalent to a reference amino acid sequence if said amino acid sequence when exposed to the immune system of a subject induces an immune reaction having a specificity of reacting with the reference amino acid sequence.
  • an antigen receptor is an antibody or B cell receptor which binds to an epitope in an antigen.
  • an antibody or B cell receptor binds to native epitopes of an antigen.
  • Increased, Induced, or Reduced As used herein, these terms or grammatically comparable comparative terms, indicate values that are relative to a comparable reference measurement.
  • an assessed value achieved with a provided pharmaceutical composition may be “increased” relative to that obtained with a comparable reference pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine).
  • an assessed value achieved in a subject may be “increased” relative to that obtained in the same subject under different conditions (e.g., prior to or after an event; or presence or absence of an event such as administration of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) as described herein, or in a different, comparable subject (e.g., in a comparable subject that differs from the subject of interest in prior exposure to a condition, e.g., absence of administration of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) as described herein.).
  • a pharmaceutical composition e.g., immunogenic composition, e.g., vaccine
  • comparative terms refer to statistically relevant differences (e.g., that are of a prevalence and/or magnitude sufficient to achieve statistical relevance). Those skilled in the art will be aware, or will readily be able to determine, in a given context, a degree and/or prevalence of difference that is required or sufficient to achieve such statistical significance.
  • the term “reduced” or equivalent terms refers to a reduction in the level of an assessed value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75% or higher, as compared to a comparable reference.
  • the term “reduced” or equivalent terms refers to a complete or essentially complete inhibition, i.e., a reduction to zero or essentially to zero.
  • the term “increased” or “induced” refers to an increase in the level of an assessed value by at least 10%, at least 20%, at least 30%, 21 11979815v1 P-627574-PC at least 40%, at least 50%, at least 80%, at least 100%, at least 200%, at least 500%, or higher, as compared to a comparable reference.
  • Ionizable refers to a compound or group or atom that is charged at a certain pH. In the context of an ionizable amino lipid, such a lipid or a function group or atom thereof bears a positive charge at a certain pH.
  • an ionizable amino lipid is positively charged at an acidic pH. In some embodiments, an ionizable amino lipid is predominately neutral at physiological pH values, e.g., in some embodiments about 7.0-7.4, but becomes positively charged at lower pH values. In some embodiments, an ionizable amino lipid may have a pKa within a range of about 5 to about 7.
  • Isolated means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not “isolated”, but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is “isolated”.
  • nucleic acid refers to a polymer of at least 10 nucleotides or more.
  • a nucleic acid is or comprises DNA.
  • a nucleic acid is or comprises RNA.
  • a nucleic acid is or comprises peptide nucleic acid (PNA).
  • PNA peptide nucleic acid
  • a nucleic acid is or comprises a single stranded nucleic acid.
  • a nucleic acid is or comprises a double- stranded nucleic acid.
  • a nucleic acid comprises both single and double- stranded portions.
  • a nucleic acid comprises a backbone that comprises one or more phosphodiester linkages.
  • a nucleic acid comprises a backbone that comprises both phosphodiester and non-phosphodiester linkages.
  • a nucleic acid may comprise a backbone that comprises one or more phosphorothioate or 5'-N-phosphoramidite linkages and/or one or more peptide bonds, e.g., as in a “peptide nucleic acid”.
  • a nucleic acid comprises one or more, or all, natural residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, a nucleic acid comprises on or more, or all, non-natural residues.
  • natural residues e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil.
  • a non-natural residue comprises a nucleoside analog (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3 - methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2- aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5 - 22 11979815v1 P-627574-PC propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 6-O-methylguanine, 2-thiocytidine, methylated bases, intercal
  • a non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) as compared to those in natural residues.
  • a nucleic acid has a nucleotide sequence that encodes a functional gene product such as an RNA or polypeptide.
  • a nucleic acid has a nucleotide sequence that comprises one or more introns.
  • a nucleic acid may be prepared by isolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, e.g., in vivo or in vitro, reproduction in a recombinant cell or system, or chemical synthesis.
  • enzymatic synthesis e.g., by polymerization based on a complementary template, e.g., in vivo or in vitro, reproduction in a recombinant cell or system, or chemical synthesis.
  • a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, or 20,000 or more residues or nucleotides long.
  • Nucleotide refers to its art-recognized meaning. When a number of nucleotides is used as an indication of size, e.g., of a polynucleotide, a certain number of nucleotides refers to the number of nucleotides on a single strand, e.g., of a polynucleotide.
  • Patient refers to any organism who is suffering or at risk of a disease or disorder or condition. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and/or humans).
  • a patient is a human. In some embodiments, a patient is suffering from or susceptible to one or more diseases or disorders or conditions. In some embodiments, a patient displays one or more symptoms of a disease or disorder or condition. In some embodiments, a patient has been diagnosed with one or more diseases or disorders or conditions. In some embodiments, a disease or disorder or condition that is amenable to provided technologies is or includes an HSV infection. In some embodiments, a patient is receiving or has received certain therapy to diagnose and/or to treat a disease, disorder, or condition. In some embodiments, a patient is a patient suffering from or susceptible to an HSV infection.
  • composition refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers.
  • active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population.
  • pharmaceutical compositions may be specially formulated for parenteral administration, for example, by subcutaneous, intramuscular, or intravenous injection as, for example, a sterile solution or suspension formulation.
  • compositions which achieves a desired reaction or a desired effect alone or together with further doses.
  • a desired reaction in some embodiments relates to inhibition of the course of the disease. In some embodiments, such inhibition may comprise slowing down the progress of a disease and/or interrupting or reversing the progress of the disease.
  • a desired reaction in a treatment of a disease may be or comprise delay or prevention of the onset of a disease or a condition.
  • compositions e.g., immunogenic compositions, e.g., vaccines
  • an effective amount of pharmaceutical compositions will depend, for example, on a disease or condition to be treated, the severity of such a disease or condition, individual parameters of the patient, including, e.g., age, physiological condition, size and weight, the duration of treatment, the type of an accompanying therapy (if present), the specific route of administration and similar factors. Accordingly, doses of pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) described herein may depend on various of such parameters. In the case that a reaction in a patient is insufficient with an initial dose, higher doses (or effectively higher doses achieved by a different, more localized route of administration) may be used.
  • Poly(A) sequence As used herein, the term “poly(A) sequence” or “poly-A tail” refers to an uninterrupted or interrupted sequence of adenylate residues which is typically located at the 3'- end of an RNA molecule. Poly(A) sequences are known to those of skill in the art and may follow the 3’-UTR in the RNAs described herein. An uninterrupted poly(A) sequence is characterized by consecutive adenylate residues. In nature, an uninterrupted poly(A) sequence is typical.
  • RNAs disclosed herein can have a poly(A) sequence attached to the free 3'-end of the RNA by a template- independent RNA polymerase after transcription or a poly(A) sequence encoded by DNA and transcribed by a template-dependent RNA polymerase. 24 11979815v1 P-627574-PC [0077]
  • Prevent refers to reducing the risk of developing the disease, disorder and/or condition and/or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time.
  • Recombinant The term “recombinant” in the context of the present disclosure means “made through genetic engineering”. In some embodiments, a “recombinant” entity such as a recombinant nucleic acid in the context of the present disclosure is not naturally occurring.
  • Reference As used herein, the term “reference” describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value. In some embodiments, a reference or control is tested and/or determined substantially simultaneously with the testing or determination of interest.
  • a reference or control is a historical reference or control, optionally embodied in a tangible medium.
  • a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment. Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and/or comparison to a particular possible reference or control.
  • Risk As will be understood from context, “risk” of a disease, disorder, and/or condition refers to a likelihood that a particular individual will develop the disease, disorder, and/or condition. In some embodiments, risk is expressed as a percentage.
  • risk is from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90 up to 100%.
  • risk is expressed as a risk relative to a risk associated with a reference sample or group of reference samples.
  • a reference sample or group of reference samples have a known risk of a disease, disorder, condition and/or event.
  • a reference sample or group of reference samples are from individuals comparable to a particular individual.
  • relative risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more.
  • risk may reflect one or more genetic attributes, e.g., which may predispose an individual toward development (or not) of a particular disease, disorder and/or condition.
  • risk may reflect one or more epigenetic events or attributes and/or one or more lifestyle or environmental events or attributes.
  • 25 11979815v1 P-627574-PC Selective or specific: The term “selective” or “specific”, when used herein in reference to an agent having an activity, is understood by those skilled in the art to mean that the agent discriminates between potential target entities, states, or cells. For example, in some embodiments, an agent is said to bind “specifically” to its target if it binds preferentially with that target in the presence of one or more competing alternative targets. In many embodiments, specific interaction is dependent upon the presence of a particular structural feature of the target entity (e.g., an epitope, a cleft, a binding site).
  • a particular structural feature of the target entity e.g., an epitope, a cleft, a binding site.
  • specificity need not be absolute. In some embodiments, specificity may be evaluated relative to that of a target-binding moiety for one or more other potential target entities (e.g., competitors). In some embodiments, specificity is evaluated relative to that of a reference specific binding moiety. In some embodiments, specificity is evaluated relative to that of a reference non-specific binding moiety.
  • Subject refers to an organism to be administered with a composition described herein, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, domestic pets, etc.) and humans.
  • a subject is a human subject. In some embodiments, a subject is suffering from a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject is susceptible to a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject displays one or more symptoms or characteristics of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject displays one or more non-specific symptoms of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject does not display any symptom or characteristic of a disease, disorder, or condition (e.g., an HSV infection).
  • a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition (e.g., an HSV infection).
  • a subject is a patient.
  • a subject is an individual to whom diagnosis and/or therapy is and/or has been administered. [0083] Suffering from: An individual who is “suffering from” a disease, disorder, and/or condition has been diagnosed with and/or displays one or more symptoms of a disease, disorder, and/or condition.
  • Susceptible to An individual who is “susceptible to” a disease, disorder, and/or condition is one who has a higher risk of developing the disease, disorder, and/or condition than does a member of the general public.
  • an individual who is susceptible to a disease, 26 11979815v1 P-627574-PC disorder and/or condition may not have been diagnosed with the disease, disorder, and/or condition.
  • an individual who is susceptible to a disease, disorder, and/or condition may exhibit symptoms of the disease, disorder, and/or condition.
  • an individual who is susceptible to a disease, disorder, and/or condition may not exhibit symptoms of the disease, disorder, and/or condition.
  • an individual who is susceptible to a disease, disorder, and/or condition will develop the disease, disorder, and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition will not develop the disease, disorder, and/or condition.
  • synthetic refers to an entity that is artificial, or that is made with human intervention, or that results from synthesis rather than naturally occurring.
  • a synthetic nucleic acid or polynucleotide refers to a nucleic acid molecule that is chemically synthesized, e.g., in some embodiments by solid-phase synthesis.
  • the term “synthetic” refers to an entity that is made outside of biological cells.
  • a synthetic nucleic acid or polynucleotide refers to a nucleic acid molecule (e.g., an RNA) that is produced by in vitro transcription using a template.
  • Therapy refers to an administration or delivery of an agent or intervention that has a therapeutic effect and/or elicits a desired biological and/or pharmacological effect (e.g., has been demonstrated to be statistically likely to have such effect when administered to a relevant population).
  • a therapeutic agent or therapy is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition.
  • a therapeutic agent or therapy is a medical intervention (e.g., surgery, radiation, phototherapy) that can be performed to alleviate, relieve, inhibit, present, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition.
  • Three prime untranslated region refers to a sequence of an RNA molecule that begins following a stop codon of a coding region of an open reading frame sequence. In some embodiments, the 3' UTR begins immediately after a stop codon of a coding region of an open reading frame sequence, e.g., in its natural context. In other embodiments, the 3' UTR does not begin immediately after stop codon of the coding region of an open reading frame sequence, e.g., in its natural context.
  • Threshold level refers to a level that are used as a reference to attain information on and/or classify the results of a measurement, for example, the results of a measurement attained in an assay.
  • a threshold level means a value measured in an assay that defines the dividing line between two subsets of a population (e.g. a batch that satisfy quality control criteria vs. a batch that does not satisfy quality control criteria).
  • a value that is equal to or higher than the threshold level defines one subset of the population, and a value that is lower than the threshold level defines the other subset of the population.
  • a threshold level can be determined based on one or more control samples or across a population of control samples. A threshold level can be determined prior to, concurrently with, or after the measurement of interest is taken. In some embodiments, a threshold level can be a range of values. [0089] Treat: As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition.
  • Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition. In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition, for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition. In some embodiments, treatment may be administered to a subject at a later-stage of disease, disorder, and/or condition.
  • Vaccination refers to the administration of a composition intended to generate an immune response, for example to a disease-associated (e.g., disease-causing) agent.
  • vaccination can be administered before, during, and/or after exposure to a disease-associated agent, and in certain embodiments, before, during, and/or shortly after exposure to the agent.
  • vaccination includes multiple administrations, appropriately spaced in time, of a vaccine composition.
  • vaccination generates an immune response to an infectious agent.
  • Vaccine refers to a composition that induces an immune response upon administration to a subject. In some embodiments, an induced immune response provides protective immunity.
  • Variant As used herein in the context of molecules, e.g., nucleic acids, proteins, or small molecules, the term “variant” refers to a molecule that shows significant structural identity with a 28 11979815v1 P-627574-PC reference molecule but differs structurally from the reference molecule, e.g., in the presence or absence or in the level of one or more chemical moieties as compared to the reference entity. In some embodiments, a variant also differs functionally from its reference molecule. In general, whether a particular molecule is properly considered to be a “variant” of a reference molecule is based on its degree of structural identity with the reference molecule.
  • any biological or chemical reference molecule has certain characteristic structural elements.
  • a variant by definition, is a distinct molecule that shares one or more such characteristic structural elements but differs in at least one aspect from the reference molecule.
  • a variant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in amino acid or nucleotide sequence and/or one or more differences in chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nucleic acid (e.g., that are attached to the polypeptide or nucleic acid backbone).
  • moieties e.g., carbohydrates, lipids, phosphate groups
  • a variant polypeptide or nucleic acid shows an overall sequence identity with a reference polypeptide or nucleic acid that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 99%.
  • a variant polypeptide or nucleic acid does not share at least one characteristic sequence element with a reference polypeptide or nucleic acid.
  • a reference polypeptide or nucleic acid has one or more biological activities.
  • a variant polypeptide or nucleic acid shares one or more of the biological activities of the reference polypeptide or nucleic acid.
  • a variant polypeptide or nucleic acid lacks one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid shows a reduced level of one or more biological activities as compared to the reference polypeptide or nucleic acid. In some embodiments, a polypeptide or nucleic acid of interest is considered to be a “variant” of a reference polypeptide or nucleic acid if it has an amino acid or nucleotide sequence that is identical to that of the reference but for a small number of sequence alterations at particular positions.
  • a variant polypeptide or nucleic acid comprises about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, or about 1 substituted residues as compared to a reference.
  • a variant polypeptide or nucleic acid comprises a very small number (e.g., fewer than about 5, about 4, about 3, about 2, or about 1) 29 11979815v1 P-627574-PC number of substituted, inserted, or deleted, functional residues (i.e., residues that participate in a particular biological activity) relative to the reference.
  • a variant polypeptide or nucleic acid comprises not more than about 5, about 4, about 3, about 2, or about 1 addition or deletion, and, in some embodiments, comprises no additions or deletions, as compared to the reference.
  • a variant polypeptide or nucleic acid comprises fewer than about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 10, about 9, about 8, about 7, about 6, and commonly fewer than about 5, about 4, about 3, or about 2 additions or deletions as compared to the reference.
  • a reference polypeptide or nucleic acid is one found in nature.
  • Vector refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
  • plasmid refers to a circular double stranded DNA loop into which additional DNA segments may be ligated.
  • vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors).
  • Other vectors e.g., non-episomal mammalian vectors
  • certain vectors are capable of directing the expression of genes to which they are operatively linked.
  • vectors are referred to herein as “expression vectors.”
  • known techniques may be used, for example, for generation or manipulation of recombinant DNA, for oligonucleotide synthesis, and for tissue culture and transformation (e.g., electroporation, lipofection).
  • Enzymatic reactions and purification techniques may be performed according to manufacturer's specifications or as commonly accomplished in the art or as described herein.
  • the foregoing techniques and procedures may be generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification. See e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual (4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
  • compositions comprising one or more RNAs (also referred to herein as “polyribonucleotides”).
  • one or more RNAs encodes a Herpes Simplex Virus (HSV) glycoprotein or immunogenic fragment thereof.
  • HSV Herpes Simplex Virus
  • an RNA is a modified RNA as described herein below.
  • an immunogenic fragment of an HSV glycoprotein comprises the ectodomain of the glycoprotein, or portion thereof.
  • an immunogenic fragment consists of the ectodomain of the glycoprotein, or portion thereof.
  • the present disclosure provides a composition comprising one or more nucleoside modified RNAs.
  • a modified RNA encodes a Herpes Simplex Virus (HSV) glycoprotein or immunogenic fragment thereof.
  • modified RNA comprises one or more pseudouridine or pseudouridine family residues.
  • an HSV glycoprotein comprises glycoprotein D (gD), glycoprotein C (gC), glycoprotein E (gE), glycoprotein B (gB), glycoprotein H (gH), glycoprotein L (gL), glycoprotein I (gI), or any combinations thereof.
  • the present disclosure provides a composition comprising one or more modified RNAs encoding HSV gD, gC, gE, gB, gH, gL, gI, or immunogenic fragments thereof.
  • the modified RNAs comprise pseudouridine-modified RNAs.
  • the present disclosure provides compositions comprising an RNA encoding HSV gD or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV gC or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV gE or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV gB or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV gH or an immunogenic fragment thereof. In 31 11979815v1 P-627574-PC another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV gL or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV gI or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV gC or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV gE or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV gC or an immunogenic fragment thereof; and (b) an RNA encoding HSV gE or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV gD or an immunogenic fragment thereof; (b) an RNA encoding HSV gC or an immunogenic fragment thereof, and (c) an RNA encoding HSV gE or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV gD or an immunogenic fragment thereof; (b) an RNA encoding HSV gC or an immunogenic fragment thereof, (c) an RNA encoding HSV gE or an immunogenic fragment thereof; and (d) an RNA encoding HSV gB or an immunogenic fragment thereof.
  • the HSV glycoproteins are HSV-2 glycoproteins or immunogenic fragments thereof. In another embodiment, the HSV glycoproteins are HSV-1 glycoproteins or immunogenic fragments thereof. In some embodiments, the HSV glycoproteins comprise both HSV-2 glycoproteins and HSV-1 glycoproteins or immunogenic fragments thereof. In another embodiment, the HSV glycoproteins comprise a mixture of HSV-2 glycoproteins and HSV-1 glycoproteins or immunogenic fragments thereof. [0107] In some embodiments, the present disclosure provides compositions comprising an RNA encoding HSV-2 gD or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-2 gC or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV-2 gE or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV-2 gE or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV-2 gB or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV-2 gH or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV-2 gL or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV-2 gI or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV-2 gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 2 gC or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV-2 gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 2 gE or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV-2 gC or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 2 gE or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV-2 gD or an immunogenic fragment thereof; (b) an RNA encoding HSV-2 gC or an immunogenic fragment thereof, and (c) an RNA encoding HSV-2 gE or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV-1 gD or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV-1 gC or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gE or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gE or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gB or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gH or an immunogenic fragment thereof.
  • the present disclosure provides compositions comprising an RNA encoding HSV-1 gL or an immunogenic fragment 33 11979815v1 P-627574-PC thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gI or an immunogenic fragment thereof. [0113] In some embodiments, the present disclosure provides a composition comprising: (a) an RNA encoding HSV-1 gD or fragment thereof; and (b) an RNA encoding HSV-1 gC or fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV-1 gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 1 gE or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV-1 gC or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 1 gE or an immunogenic fragment thereof.
  • the present disclosure provides a composition comprising: (a) an RNA encoding HSV-1 gD or an immunogenic fragment thereof; (b) an RNA encoding HSV-1 gC or an immunogenic fragment thereof, and (c) an RNA encoding HSV-1 gE or an immunogenic fragment thereof.
  • any of the compositions as described herein consists essentially of one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof.
  • any of the compositions as described herein consists of one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof.
  • compositions comprising an RNA encoding an HSV gD protein or immunogenic fragment thereof, an RNA encoding an HSV gC protein or immunogenic fragment thereof, an RNA encoding an HSV gE protein or immunogenic fragment thereof, and RNAs encoding one or more additional HSV glycoproteins.
  • said additional HSV glycoproteins comprise gB or immunogenic fragment thereof, gH or immunogenic fragment thereof, gL or immunogenic fragment thereof, gI or immunogenic fragment thereof, or any combination thereof.
  • compositions of the present disclosure and for use in the methods of the present disclosure comprise both HSV-2 glycoproteins or glycoprotein fragments and HSV- 1 glycoproteins or glycoprotein fragments.
  • compositions of the present 34 11979815v1 P-627574-PC disclosure and for use in the methods of the present disclosure comprise a mixture of HSV-2 glycoproteins or glycoprotein fragments and HSV-1 glycoproteins or glycoprotein fragments.
  • a composition of the present disclosure comprises HSV-2 gC, HSV-1 gD, and HSV-2 gE, or immunogenic fragments thereof.
  • a composition of the present disclosure comprises HSV-1 gC, HSV-2 gD, and HSV-2 gE, or immunogenic fragments thereof.
  • a composition of the present disclosure comprises HSV-2 gC, HSV-2 gD, and HSV-1 gE, or immunogenic fragments thereof.
  • a composition of the present disclosure comprises HSV-1 gC, HSV-1 gD, and HSV- 2 gE, or immunogenic fragments thereof.
  • composition of the present disclosure comprises HSV-1 gC, HSV-2 gD, and HSV-1 gE, or immunogenic fragments thereof.
  • a composition of the present disclosure comprises HSV-2 gC, HSV-1 gD, and HSV-1 gE, or immunogenic fragments thereof.
  • the compositions of the present disclosure comprise one or more additional (i) HSV-1 glycoproteins or immunogenic fragments thereof, (ii) HSV-2 glycoproteins or immunogenic fragments thereof, or (iii) both HSV-1 or immunogenic fragments thereof and HSV-2 glycoproteins or immunogenic fragments thereof, as described herein.
  • a composition of the present disclosure comprising HSV-2 gC, HSV-1 gD, and HSV-2 gE may further comprise HSV-1 gI.
  • a composition of the present disclosure comprising HSV-2 gC, HSV-2 gD, and HSV-2 gE may further comprise HSV- 1 gB.
  • encoding refers to an RNA molecule that contains a gene that encodes a protein of interest or a fragment thereof.
  • an RNA molecule comprises a coding sequence that encodes a protein of interest or a fragment thereof.
  • one or more other proteins or fragments thereof is also encoded.
  • the protein of interest or a fragment thereof is the only protein encoded.
  • Immunogenic fragment refers to a portion of a protein that is immunogenic and elicits a protective immune response when administered to a subject.
  • Immunogenicity or “immunogenic” is used herein to refer to the innate ability of a protein, peptide, protein fragment, nucleic acid, antigen or organism to elicit an immune response in an animal when the protein, peptide, nucleic acid, antigen or organism is administered to the 35 11979815v1 P-627574-PC animal.
  • enhancing the immunogenicity refers to increasing the ability of a protein, peptide, protein fragment, nucleic acid, antigen or organism to elicit an immune response in an animal when the protein, peptide, protein fragment, nucleic acid, antigen or organism is administered to an animal.
  • the increased ability of a protein, peptide, protein fragment, nucleic acid, antigen or organism to elicit an immune response can be measured by, in some embodiments, a greater number of antibodies to a protein, peptide, protein fragment, nucleic acid, antigen or organism, a greater diversity of antibodies to an antigen or organism, a greater number of T-cells specific for a protein, peptide, protein fragment, nucleic acid, antigen or organism, a greater cytotoxic or helper T-cell response to a protein, peptide, protein fragment, nucleic acid, antigen or organism, and the like.
  • a protein, peptide, protein fragment, nucleic acid or organism can be antigenic.
  • Antigenic refers to a protein, peptide, protein fragment, nucleic acid, or organism capable of specifically interacting with an antigen recognition molecule of the immune system, e.g., an immunoglobulin (antibody) or T cell antigen receptor.
  • An antigenic protein, peptide, protein fragment contains, in another embodiment, an epitope of at least about 8 amino acids (AA).
  • An antigenic portion of a protein, peptide, protein fragment, nucleic acid, or organism also called herein an “epitope” can be a portion that is immunodominant for antibody or T cell receptor recognition, or it can be a portion used to generate an antibody to the molecule by conjugating an antigenic portion to a carrier (e.g., a carrier polypeptide) for immunization.
  • a carrier e.g., a carrier polypeptide
  • a molecule that is antigenic need not be itself immunogenic, i.e., capable of eliciting an immune response without a carrier.
  • “Functional” is used herein to refer to the innate ability of a protein, peptide, nucleic acid, fragment or a variant thereof to exhibit a biological activity or function.
  • such a biological function is its binding property to an interaction partner, e.g., a membrane- associated receptor, and in another embodiment, its trimerization property.
  • these biological functions may in fact be changed, e.g., with respect to their specificity or selectivity, but with retention of the basic biological function.
  • fragment is used herein to refer to a protein or polypeptide that is shorter or comprises fewer amino acids than the full length protein or polypeptide.
  • fragment refers to a nucleic acid encoding the protein fragment that is shorter or comprises fewer nucleotides than the full length nucleic acid.
  • the fragment is an N-terminal fragment. In another embodiment, the fragment is a C-terminal fragment. In some embodiments, the fragment is an intrasequential section of the protein, peptide, or nucleic acid. In another embodiment, the fragment is an immunogenic intrasequential section of the protein, peptide or nucleic acid. In another embodiment, the fragment is a functional intrasequential section within the protein, peptide or nucleic acid. In another embodiment, the fragment is an N-terminal immunogenic fragment. In some embodiments, the fragment is a C-terminal immunogenic fragment. In another embodiment, the fragment is an N- terminal functional fragment.
  • an “immunogenic fragment” of a protein as described in the present disclosure refers to a portion of the protein that is immunogenic, in one embodiment and in another embodiment, elicits a protective immune response when administered to a subject.
  • compositions comprising RNAs, wherein each of said RNAs encodes a) HSV glycoprotein D (gD) or an immunogenic fragment thereof, b) HSV glycoprotein C (gC) or an immunogenic fragment thereof, c) HSV glycoprotein E (gE) or an immunogenic fragment thereof, or any combination thereof.
  • the present disclosure provides a composition comprising an RNA encoding an HSV gD or an immunogenic fragment thereof, an RNA encoding an HSV gC or an immunogenic fragment thereof, and an RNA encoding an HSV gE or an immunogenic fragment thereof.
  • compositions of RNA encoding HSV-1 gD (gD1) or an immunogenic fragment thereof are protective against HSV-1 infection.
  • combination compositions of RNA encoding HSV gC (gC1) or an immunogenic fragment thereof /HSV-1 gD (gD1) or an immunogenic fragment thereof /HSV-1 gE (gE1) or an immunogenic fragment thereof can are protective against HSV-1.
  • compositions of RNA encoding gD2 or an immunogenic fragment thereof can be protective against HSV-2 infection ( Figures 7-10).
  • compositions of RNA encoding gC2 or an immunogenic fragment thereof /gD2 or an immunogenic fragment thereof /gE2 or an immunogenic fragment thereof can confer superior protection compared with compositions containing RNA encoding gC2 alone, gD2 alone, or gE2 alone.
  • 37 11979815v1 P-627574-PC [0131]
  • inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure can increase the efficaciousness of anti-gD antibodies elicited by the composition.
  • inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure can increase the dose of RNA encoding gD or an immunogenic fragment thereof required to elicit antibodies that inhibit binding of gD to a cellular receptor.
  • inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure can decrease the dose of RNA encoding gD or an immunogenic fragment thereof required to elicit antibodies that inhibit binding of gD or an immunogenic fragment thereof to a cellular receptor when a dose of RNA encoding a gD or an immunogenic fragment thereof is administered separately from RNAs encoding the gC protein, gE protein or immunogenic fragments thereof.
  • RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure enhances the effectiveness of an innate immune response.
  • the innate immune response is an antibody-mediated immune response.
  • the innate immune response is a non-antibody-mediated immune response.
  • the innate immune response is an NK (natural killer) cell response.
  • the innate immune response is any other innate immune response known in the art.
  • inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure increases the efficaciousness of antibodies elicited by the composition against one of the above glycoproteins.
  • inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure decreases the dose of one of the above glycoproteins required to elicit antibodies that inhibit binding of the glycoprotein to a cellular receptor thereof, when a dose of one of the glycoproteins is administered separately from one of the other glycoproteins.
  • a composition comprises (i) one or more RNAs encoding HSV glycoproteins or immunogenic fragments thereof and (ii) lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes.
  • PLX polyplexes
  • LPLX lipidated polyplexes
  • Glycoprotein D HSV-1 gD [0135]
  • a composition of the present disclosure comprises an RNA encoding HSV-1 gD protein.
  • a composition comprises an RNA encoding a fragment of an HSV-1 gD protein (e.g., an immunogenic fragment).
  • a nucleotide sequence of the RNA encoding an HSV-1 gD fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCU GCUGCUGCUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCAAGUACGC CCUGGCCGACGCCUCCCUGAAGAUGGCCGACCCCAACCGCUUCCGCGGCAAGG ACCUGCCCGUGCUGGACCAGCUGACCGACCCCCCCGGCGUGCGCCGCGUGUAC CACAUCCAGGCCGGCCUGCCCGACCCCUUCCAGCCCCCCUCCCUGCCCAUCACC GUACUACGCCGUGCUGGAGCGCGCCGCUCCGUGCCCACCAUCACC GUACUACGCCGUGCUGGAGCGCCCCC
  • underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243).
  • bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 19) to assist expression of the HSV-1 gD fragment.
  • italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264).
  • a nucleotide sequence of the RNA encoding an HSV-1 gD fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or any combinations thereof.
  • an HSV-1 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-331 of gD (e.g., from HSV- 1 Patton strain), as set forth in the following amino acid sequence: KYALADASLKMADPNRFRGKDLPVLDQLTDPPGVRRVYHIQAGLPDPFQPPSLPITV YYAVLERACRSVLLNAPSEAPQIVRGASEDVRKQPYNLTIAWFRMGGNCAIPITVME YTECSYNKSLGACPIRTQPRWNYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKIN DWTEITQFILEHRAKGSCKYALPLRIPPSACLSPQAYQQGVTVDSIGMLPRFIPENQRT VAVYSLKIAGWHGPKAPYTSTLLPPELSETPNATQPELAPEDPEDS
  • an HSV-1 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 2.
  • the HSV-1 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-333 of gD (e.g., from HSV- 1 Patton strain), as set forth in the following amino acid sequence: KYALADASLKMADPNRFRGKDLPVLDQLTDPPGVRRVYHIQAGLPDPFQPPSLPITV YYAVLERACRSVLLNAPSEAPQIVRGASEDVRKQPYNLTIAWFRMGGNCAIPITVME 40 11979815v1 P-627574-PC YTECSYNKSLGACPIRTQPRWNYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKIN DWTEITQFILEHRAKGSCKYALPLRIPPSACLSPQAYQQGVTVDSIGMLPRFIPENQRT VAVYSLKIAGWHGPK
  • an HSV-1 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 134.
  • full length HSV-1 gD encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MGGAAARLGAVILFVVIVGLHGVRGKYALADASLKLADPNRFRRKDLPVLDQLTDP PGVRRVYHIQAGLPDPFQPPSLPITVYYAVLERACRSVLLNAPSEAPQIVRGASEDVR KQPYNLTIAWFRMGGNCAIPITVMEYTECSYNKSLGACPIRTQPRWNYYDSFSAVSE DNLGFLMHAPAFETAGTYLRLVKINDWTEITQFILEHRAKGSCKYALPLRIPPSACLS PQAYQQGVTVDSIGMLPRFIPENQRTVAVYSLKIAGWHGPKAPYTSTLLPPELSETPN ATQPELAPEAPEDSALLEDPVGTVAPQIPPN
  • an HSV-1 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 3.
  • an HSV-1 gD, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any one of the following GenBank Accession Numbers: AAL90884.1 (KHS2 strain), AAL90883.1 (KHS1 strain), AAK93950.1 (F strain), AAB59754.1 (F strain), AAA19631.1 (mutant strain not identified), AAA19630.1 (mutant strain not identified), or AAA19629.1 (strain not identified).
  • an HSV-1 gD, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid 41 11979815v1 P-627574-PC sequences as set forth in any of the following GenBank Accession Numbers: A1Z0Q5.2, AAA45780.1, AAA45785.1, AAA45786.1, AAA96682.1, AAK19597.1, AAN74642.1, ABI63524.1, ABM52978.1, ABM52979.1, ABM52980.1, ABM52981.1, ABM66847.1, ABM66848.1, ACM62295.1, ADD60053.1, ADD60130.1, ADM22389.1, ADM22466.1, ADM22542.1, ADM22619.1, ADM22696.1, ADM22773.1, ADM22849.1, ADM22926.1, ADM23003.1, ADM23079.1, ADM23155.1, ADM23231.1, ADM23309.1, ADM23383.1, A
  • a composition comprises an RNA encoding an HSV-2 gD protein.
  • a composition comprises an RNA encoding a fragment of an HSV-2 gD protein (e.g., an immunogenic fragment).
  • a nucleotide sequence of the RNA encoding an HSV-2 gD fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAA GCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAGCAAU UUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGACCCGCCUGACCGUGCU GGCCCUGCUGGCCGGCCUGCUGGCCUCCUCCCGCGCCAAGUACGCCCUGGCCG ACCCCUCCCUGAAGAUGGCCGACCCCAACCGCUUCCGCGGCAAGAACCUGCCCGU GCUGGACCAGCUGACCGACCCCCCCGGCGUGAAGCGCGUGUACCACAUCCAGCCC UCCCUGGAGGACCGACCGACCCCCCCGGCGUGAAGCGCGUGUACCACAUCCAGCCC UCCCUGGAGGACCGACCGACCCCCCCGGCGUGAAGCGCGUGUACCACAUCCAGCCC UCCCUG
  • underlined residues represent 5’ untranslated sequence (SEQ ID NO: 243).
  • bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 20) to assist expression of the HSV-2 gD fragment.
  • italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264).
  • a nucleotide sequence of the RNA encoding an HSV-2 gD fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or any combinations thereof.
  • an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-331 of gD (e.g., from HSV- 2 strain 333 or US6), as set forth in the following amino acid sequence: KYALADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVY YAVLERACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVME YTECPYNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKIN DWTEITQFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQR TVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDS
  • an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 5.
  • an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 5.
  • an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 30-331 of gD from HSV-2 (e.g., strain 333 or US6), as set forth in the following amino acid sequence: ADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVL ERACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECP 44 11979815v1 P-627574-PC YNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEI TQFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALY SLKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPN WHIPSIQDVAPHH
  • an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 234.
  • an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 234.
  • an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 31-331 of gD from HSV-2 (e.g., strain 333 or US6), as set forth in the following amino acid sequence: DPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLE RACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECPY NKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEIT QFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALYS LKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPN WHIPSIQDVAPHH (SEQ ID NO: 110).
  • an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 110.
  • an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 110.
  • an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-333 of gD from HSV-2 (e.g., strain 333 or US6), as set forth in the following amino acid sequence: DPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLE RACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECPY NKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEIT QFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALYS LKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPN WHIPSIQDVAPHHAP (SEQ ID NO: 120).
  • an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 120.
  • an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 120.
  • the full length HSV-2 gD encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: [0160] MGRLTSGVGTAALLVVAVGLRVVCAKYALADPSLKMADPNRFRGKNLPVLDQ LTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLERACRSVLLHAPSEAPQIVRGASDEA RKHTYNLTIAWYRMGDNCAIPITVMEYTECPYNKSLGVCPIRTQPRWSYYDSFSAVSED NLGFLMHAPAFETAGTYLRLVKINDWTEITQFILEHRARASCKYALPLRIPPAACLTSKA YQQGVTVDSIGMLPRFIPENQRTVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQPE LVPEDPEDSALLEDPAGTVSSQIPPNWHIPSIQDVAPHHAPAAPSNPGLIIGALAGSTLAV LVIGGIAFWVRRRAQMAPKRLRLPHIR
  • an HSV-2 gC comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 6. In some embodiments, an HSV-2 gC has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 6.
  • an HSV-2 gD, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in GenBank Accession Numbers: 1003204A, AAA45841.1, AAA45842.1, AAB60552.1, AAB60553.1, AAB60554.1, AAB60555.1, AAB72102.1, AAS01730.1, AAW23130.1, AAW23131.1, AAW23132.1, AAW23133.1, AAW23134.1, ABS84899.1, ABU45433.1, ABU45434.1, ABU45435.1, ABU45461.1, ABU45462.1, ACA28831.1, AEV91405.1, AFM93876.1, AFS18198.1, AFS18199.1, AFS18200.1, AFS18201.1, AFS18202.1, AFS18203.1, AFS18204.1, AFS18205.1, AFS18206.1, AFS18207.1, AFS18208.1,
  • the gD protein or fragment includes Y63.
  • the gD protein or fragment includes R159.
  • the gD protein or fragment includes D240.
  • the gD protein or fragment includes P246.
  • the gD protein or fragment includes a residue selected from Y63, R159, D240, and P246. In another embodiment, inclusion of one of these residues elicits antibodies that inhibit binding to nectin-1.
  • gD amino acid residues includes the residues of the signal peptide encoded by the signal sequence. Thus, residue one of the mature protein is referred to as “26.”
  • Each RNA encoding HSV-1 gD and HSV-2 gD protein or immunogenic fragment thereof represents a separate embodiment of the present disclosure. 47 11979815v1 P-627574-PC
  • an HSV gD, gC, and gE proteins, or immunogenic fragments thereof, encoded by the modified RNA as disclosed herein are described in US Patent Publication No.2013-0028925-A1, which is incorporated by reference herein in its entirety.
  • a gD protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure is an immunogenic fragment.
  • a gD immunoprotective antigen need not be the entire protein.
  • the protective immune response generally involves, in another embodiment, an antibody response.
  • mutants, sequence conservative variants, and functional conservative variants of gD are useful in methods and compositions of the present disclosure, provided that all such variants retain the required immuno-protective effect.
  • the immunogenic fragment can comprise an immuno-protective gD antigen from any strain of HSV.
  • the immunogenic fragment can comprise sequence variants of HSV, as found in infected individuals.
  • a composition of the present disclosure comprises an RNA encoding HSV-1 gC protein.
  • a composition comprises an RNA encoding a fragment of an HSV-1 gC protein (e.g., an immunogenic fragment).
  • a nucleotide sequence of the RNA encoding an HSV-1 gC fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGGCCAUCUCCGG CGUGCCCGUGCUGGGCUUCUUCAUCAUCGCCGUGCUGAUGUCCGCCCAGG AGUCCUGGGCCGAGACCGCCUCCACCGGCCCCACCAUCACCGCCGGCCGUG ACCAACGCCUCCGAGGCCCCCACCUCCGGCUCCCGGCUCCGCCGCCUCCCCC GAGGUGACCCCCACCUCCACCCCCAACCCCAACAACGUGACCCAGAACAAGAC CACCCCCACCGAGCCCGCCUCCCCCCACCACCCCCAAGCCCACCUCCACCCC CAAGUCCCCCCCCCCCC GAGGUGACCCCCACCUCC
  • underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243).
  • bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 21) to assist expression of the HSV-1 gC fragment.
  • italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264).
  • a nucleotide sequence of the RNA encoding an HSV-1 gC fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or any combinations thereof.
  • an HSV-1 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 27-457 of gC from HSV-1 (e.g., KOS strain), as set forth in the following amino acid sequence: ETASTGPTITAGAVTNASEAPTSGSPGSAASPEVTPTSTPNPNNVTQNKTTPTEPASPP TTPKPTSTPKSPPTSTPDPKPKNNTTPAKSGRPTKPPGPVWCDRRDPLARYGSRVQIR CRFRNSTRMEFRLQIWRYSMGPSPPIAPAPDLEEVLTNITAPPGGLLVYDSAPNLTDP HVLWAEGAGPGADPPLYSVTGPLPTQRLIIGEVTPATQGMYYLAWGRMDSPHEYGT WVRVRMFRPPSLTLQPHAVMEGQPFKATCTAAAYYY
  • an HSV-1 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 8.
  • an HSV-1 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 8.
  • the HSV-1 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 25-457 of gC from HSV-1 (e.g., KOS strain), as set forth in the following amino acid sequence: GSETASTGPTITAGAVTNASEAPTSGSPGSAASPEVTPTSTPNPNNVTQNKTTPTEPAS PPTTPKPTSTPKSPPTSTPDPKPKNNTTPAKSGRPTKPPGPVWCDRRDPLARYGSRVQ IRCRFRNSTRMEFRLQIWRYSMGPSPPIAPAPDLEEVLTNITAPPGGLLVYDSAPNLTD PHVLWAEGAGPGADPPLYSVTGPLPTQRLIIGEVTPATQGMYYLAWGRMDSPHEYG TWVRVRMFRPPSLTLQPHAVMEGQPFKATCTAAAYYPRNPVEFDWFEDDRQVFNP GQIDTQTHEHPDGFTTVSTVTSEAVGGQVPPRTFTCQMTWHRDSVTFSR
  • an HSV-1 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 240.
  • an HSV-1 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 240. 50 11979815v1 P-627574-PC
  • a gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 27-457 of gC from an HSV-1 strain.
  • the full length HSV-1 gC encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MAPGRVGLAVVLWGLLWLGAGVAGGSETASTGPTITAGAVTNASEAPTSGSPGSA ASPEVTPTSTPNPNNVTQNKTTPTEPASPPTTPKPTSTPKSPPTSTPDPKPKNNTTPAKS GRPTKPPGPVWCDRRDPLARYGSRVQIRCRFRNSTRMEFRLQIWRYSMGPSPPIAPA PDLEEVLTNITAPPGGLLVYDSAPNLTDPHVLWAEGAGPGADPPLYSVTGPLPTQRLI IGEVTPATQGMYYLAWGRMDSPHEYGTWVRVRMFRPPSLTLQPHAVMEGQPFKAT CTAAAYYPRNPVEFDWFEDDRQVFNPGQIDTQTHEHPDGFTTVSTVTSEAVGGQVP PRTFTCQMTWHRDSVTFSRRNATGLALVLPRPTITMEFGVRH
  • an HSV-2 gC comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 9.
  • an HSV-2 gC has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 9.
  • an HSV-1 gC encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: AAA45779.1, AAA96680.1, ABI63505.1, ABM52973.1, ABM52976.1, ABM52977.1, ACM62267.1, ADD60042.1, ADD60119.1, ADM22367.1, ADM22444.1, ADM22520.1, ADM22597.1, ADM22674.1, ADM22751.1, ADM22827.1, ADM22904.1, ADM22981.1, ADM23057.1, ADM23133.1, ADM23210.1, ADM23287.1, ADM23361.1, ADM23435.1, ADM23509.1, ADM23583.1, ADM23658.1, ADM23733.1, ADM23809.1, AEQ77075.1, AEQ77099.1, AER37628.1, AER37697.1, AER37767.1, AER37
  • a composition comprises an RNA encoding an HSV-2 gC protein.
  • a composition comprises an RNA encoding a fragment of an HSV-2 gC protein (e.g., immunogenic fragment).
  • a nucleotide sequence of the RNA encoding an HSV-2 gC fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCU GCUGCUGCUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCGCCUCCCC CGGCCGCACCAUCACCGUGGGCCCCCGCGGCAACGCCUCCAACGCCGCCCCCUC CGCCUCCCCCCGCAACGCCUCCGCCCCCCGCACCACCCCCACCCCCCCCCAGCCC CGCAAGGCCACCAAGUCCACCAAGUCCCCCCCCCAAG ACCGGCCCCCCCAAGACCUCCUCCGAGCCCGUGCUGCAACCGCCACGACCCC CUGGGGCCACCAAGUCCCCCCCCACCAGCCCGCCCCCCCAAG
  • all uridine residues are 1-methyl-pseudouridine.
  • underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243).
  • bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 19) to assist expression of the HSV-2 gC fragment.
  • italicized residues represent 3’ untranslated sequences and a poly adenylation tail (SEQ ID NO: 244).
  • a nucleotide sequence of the RNA encoding an HSV-2 gC fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or any combinations thereof.
  • the sequence of the HSV-2 gC fragment is as set forth in SEQ ID NO: 25.
  • an HSV-2 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 27-426 of gC from HSV-2 (e.g.
  • strain 333 or UL44 as set forth in the following amino acid sequence: ASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKT GPPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTESRLQIWRYATATDAEIGTAP SLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRL IIEELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATC TAATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGPP RTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFA WFLGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEH H (SEQ ID NO: 11).
  • an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 11.
  • an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 11.
  • an HSV-2 gC fragment comprises the following amino acid sequence: ASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKT GPPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTEFRLQIWRYATATDAEIGTAP SLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRL IIEELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATC TAATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGPP RTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFA WFLGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIP
  • an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 28.
  • an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 28.
  • the HSV-2 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 28-426 of gC from HSV-2 (e.g., strain 333 or UL44), as set forth in the following amino acid sequence: SPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTG PPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTESRLQIWRYATATDAEIGTAPS LEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLII EELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCT AATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGPPR TFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPE
  • an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 223.
  • an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 223.
  • an HSV-2 gC fragment comprises the following amino acid sequence: SPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTG PPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTEFRLQIWRYATATDAEIGTAPS LEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLII EELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCT AATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGPPR TFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFAWF LGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGI
  • an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 224.
  • an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 224.
  • the HSV-2 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-426 of gC from HSV-2 (e.g., strain 333 or UL44), as set forth in the following amino acid sequence: SASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPK TGPPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTESRLQIWRYATATDAEIGTA PSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQR LIIEELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKAT CTAATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGP PRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVT
  • an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 241.
  • an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 241.
  • an HSV-2 gC fragment comprises the following amino acid sequence: SASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPK TGPPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTEFRLQIWRYATATDAEIGTA PSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQR LIIEELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKAT CTAATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGP PRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFA WFLGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPV
  • an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 242.
  • an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 242.
  • the full length HSV-2 gC encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MALGRVGLAVGLWGLLWVGVVVLANASPGRTITVGPRGNASNAAPSASPRNASA PRTTPTPPQPRKATKSKASTAKPAPPPKTGPPKTSSEPVRCNRHDPLARYGSRVQIRC RFPNSTRTEFRLQIWRYATATDAEIGTAPSLEEVMVNVSAPPGGQLVYDSAPNRTDP HVIWAEGAGPGASPRLYSVVGPLGRQRLIIEELTLETQGMYYWVWGRTDRPSAYGT WVRVRVFRPPSLTIHPHAVLEGQPFKATCTAATYYPGNRAEFVWFEDGRRVFDPAQI HTQTQENPDGFSTVSTVTSAAVGGQGPPRTFTCQLTWHRDSVSFSRRNASGTASVLP RPTITMEFTGDHAVCTAGCVPEGVTFAWFLGDDSSPAEKVASQ
  • an HSV-2 gC comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 12. In some embodiments, an HSV-2 gC has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 12.
  • an HSV-2 gC encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: AAA20532.1, AAA66442.1, AAB60549.1, AAB60550.1, AAB60551.1, AAB72101.1, ABU45429.1, ABU45430.1, ABU45431.1, ABU45432.1, ABU45459.1, ABU45460.1, AEV91348.1, AEV91383.1, AEV91407.1, AFM93864.1, AHG54708.1, AKC42808.1, AKC59285.1, AKC59357.1, AKC59428.1, AKC59499.1, AKC59570.1, AMB66008.1, AMB66079.1, AMB66151.1, AMB66224.1, AMB66252.1, AMB66253.1, AMB66368.1, AMB66441.1, AQZ5573
  • an HSV gC protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises a properdin interfering domain
  • “Properdin-interfering domain” refers, in some embodiments, to a domain that blocks or inhibits binding of a host C3b molecule with a host properdin molecule. In another embodiment, the term refers to a domain that blocks or inhibits an interaction of a host C3b molecule with a host properdin molecule.
  • an HSV gC protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure is a C5 interfering domain. In another embodiment, the gC protein fragment is a portion of a C5 interfering domain.
  • C5-interfering domain refers, in another embodiment, to a domain that interferes with binding of a host C3b molecule with a host C5 molecule. In another embodiment, the term refers to a domain that interferes with the interaction of a host C3b molecule with a host C5 molecule.
  • 58 11979815v1 P-627574-PC Each RNA encoding HSV-1 gCor HSV-2 gC protein or fragment thereof represents a separate embodiment of the present disclosure.
  • a gC protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure is an immunogenic fragment.
  • a gC immunoprotective antigen need not be the entire protein.
  • the protective immune response generally involves, in another embodiment, an antibody response.
  • mutants, sequence conservative variants, and functional conservative variants of gC are useful in methods and compositions of the present disclosure, provided that all such variants retain the required immuno-protective effect.
  • the immunogenic fragment can comprise an immuno-protective gC antigen from any strain of HSV.
  • the immunogenic fragment can comprise sequence variants of HSV, as found in infected individuals.
  • Glycoprotein E HSV-1 gE [0203]
  • a composition of the present disclosure comprises an RNA encoding HSV-1 gE protein.
  • a composition comprises an RNA encoding a fragment of an HSV-1 gE protein (e.g., immunogenic fragment).
  • a nucleotide sequence of the RNA encoding an HSV-1 gE fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCU GCUGCUGCUGAUCGCCCUGGCCCUGGUGACCAACUCCAAGACCUC CUGGCGCCGCGUGUCCGUGGGCGAGGACGUGUCCCUGCUGCCCGCCCCCGGCC CCACCGGCCGCGGCCACCCAGAAGCUGCUGUGGGCCGUGGAGCCCCUGGAC GGCUGCGGCCCCCUGCACCCCUCCUGGGUGUCCCUGAUGCCCCCCAAGCAGGU GCCCGAGACCGUGGUGGACGCCGCCUGCUGUGGGCCGUGGAGCCCCUGGAC GGCUGCGGCCCCCUGCACCCCUCCUGG
  • all uridine residues are 1-methyl-pseudouridine.
  • underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243).
  • bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 19) to assist expression of the HSV-1 gE fragment.
  • italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264).
  • a nucleotide sequence of the RNA encoding an HSV-1 gE fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof.
  • the sequence of the HSV-1 gE fragment is as set forth in SEQ ID NO: 26.
  • an HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-409 of gE from HSV-1 (e.g., NS strain), as set forth in the following amino acid sequence: KTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLWAVEPLDGCGPLHPSWVSLMPPKQV PETVVDAACMRAPVPLAMAYAPPAPSATGGLRTDFVWQERAAVVNRSLVIYGVRE TDSGLYTLSVGDIKDPARQVASVVLVVQPAPVPTPPPTPADYDEDDNDEGEGEDESL 60 11979815v1 P-627574-PC AGTPASGTPRLPPSPAPPRSWPSAPEVSHVRGVTVRMETPEAILFSPGEAFSTNVSIHA IAHDDQTYTMDVVWLRFDVPTSCAEMRIYESCLYHPQLPECLSPADAPCAASTWTS RLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLAWQA
  • an HSV-1 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 14.
  • an HSV-1 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 14.
  • a gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-409 of gE from an HSV-1 strain (SEQ ID NO: 14).
  • an HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 21-409 of gE from HSV-1 (e.g., NS strain or US8), as set forth in the following amino acid sequence: GTPKTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLWAVEPLDGCGPLHPSWVSLMPP KQVPETVVDAACMRAPVPLAMAYAPPAPSATGGLRTDFVWQERAAVVNRSLVIYG VRETDSGLYTLSVGDIKDPARQVASVVLVVQPAPVPTPPPTPADYDEDDNDEGEGE DESLAGTPASGTPRLPPSPAPPRSWPSAPEVSHVRGVTVRMETPEAILFSPGEAFSTNV SIHAIAHDDQTYTMDVVWLRFDVPTSCAEMRIYESCLYHPQLPECLSPADAPCAAST WTSRLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLAWQAASVNLEFRDASPQHSGLY L
  • an HSV-1 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 248.
  • an HSV-1 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 248.
  • an HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 21-409 of gE from an HSV-1 strain.
  • an HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-309 of gE from HSV-1 (e.g., NS strain or US8), as set forth in the following amino acid sequence: GTPKTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLWAVEPLDGCGPLHPSWVSLMPP KQVPETVVDAACMRAPVPLAMAYAPPAPSATGGLRTDFVWQERAAVVNRSLVIYG VRETDSGLYTLSVGDIKDPARQVASVVLVVQPAPVPTPPPTPADYDEDDNDEGEGE DESLAGTPASGTPRLPPSPAPPRSWPSAPEVSHVRGVTVRMETPEAILFSPGEAFSTNV SIHAIAHDDQTYTMDVVWLRFDVPTSCAEMRIYESCLYHPQLPECLSPADAPCAAST WTSRLAVRSYAGCSRTNPPPRCSAEAHMEPFPG
  • an HSV-1 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 248.
  • an HSV-1 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 248.
  • a full length HSV-1 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MDRGAVVGFLLGVCVVSCLAGTPKTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLW AVEPLDGCGPLHPSWVSLMPPKQVPETVVDAACMRAPVPLAMAYAPPAPSATGGL RTDFVWQERAAVVNRSLVIYGVRETDSGLYTLSVGDIKDPARQVASVVLVVQPAPV PTPPPTPADYDEDDNDEGEGEDESLAGTPASGTPRLPPSPAPPRSWPSAPEVSHVRGV TVRMETPEAILFSPGEAFSTNVSIHAIAHDDQTYTMDVVWLRFDVPTSCAEMRIYES CLYHPQLPECLSPADAPCAASTWTSRLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLA WQAASVNLEFRDASPQHSGLYLCVVYVNDHIHAWGHITINTAAQYRNAV
  • an HSV-1 gE comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ 62 11979815v1 P-627574-PC ID NO: 15.
  • an HSV-1 gE has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO:15.
  • an HSV-1 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: AAA45779.1, AAA96680.1, ABI63526.1, ACM62297.1, ADD60055.1, ADD60132.1, ADM22391.1, ADM22468.1, ADM22544.1, ADM22621.1, ADM22698.1, ADM22775.1, ADM22851.1, ADM22928.1, ADM23005.1, ADM23081.1, ADM23157.1, ADM23233.1, ADM23311.1, ADM23385.1, ADM23459.1, ADM23533.1, ADM23607.1, ADM23682.1, ADM23757.1, ADM23833.1, ADN34689.1, ADN34692.1, ADN34695.1, AEQ77099.1, AER37649.1, AER37717.1, AER37788.1, AER37859.1, AER
  • a composition comprises an RNA encoding an HSV-2 gE protein. In another embodiment, a composition comprises an RNA encoding a fragment of an HSV-2 gE protein (e.g., immunogenic fragment).
  • a nucleotide sequence of the RNA encoding an HSV-2 gE fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCU GCUGCUGCUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCCGCACCUC CUGGAAGCGCGUGACCUCCGGCGAGGACGUGGUGCUGCUGCCCGCCCCCGCCG GCCCCGAGGAGCGCACCCGCCCACAAGCUGCUGUGGGCCGCCGAGCCCCUG GACGCCUGCGGCCCCCUGCGCCCCUCCUGGGUGGCCCUGUGGCCCCCGCCGC GUGCUGGAGACCGUGGUGGACGCCGCCUGCCCCCCCCGCCGC GUGCUGGAGACCGUGGUGGACGCCGCCUGCCCCCCG
  • underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243).
  • bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 19) to assist expression of the HSV-2 gE fragment.
  • italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264).
  • a nucleotide sequence of the RNA encoding an HSV-2 gE fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof.
  • an HSV-2 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-405 of gE from HSV-2 (e.g., strain 2.12 or US8) as set forth in the following amino acid sequence: RTSWKRVTSGEDVVLLPAPAGPEERTRAHKLLWAAEPLDACGPLRPSWVALWPPR RVLETVVDAACMRAPEPLAIAYSPPFPAGDEGLYSELAWRDRVAVVNESLVIYGAL ETDSGLYTLSVVGLSDEARQVASVVLVVEPAPVPTPTPDDYDEEDDAGVSERTPVSV 65 11979815v1 P-627574-PC PPPTPPRRPPVAPPTHPRVIPEVSHVRGVTVHMETPEAILFAPGETFGTNVSIHAIAHD DGPYAMDVVWMRFDVPSSCAEMRIYEACLYHPQLPEC
  • an HSV-2 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 17.
  • an HSV-2 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 17.
  • full length HSV-2 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MARGAGLVFFVGVWVVSCLAAAPRTSWKRVTSGEDVVLLPAPAERTRAHKLLWA AEPLDACGPLRPSWVALWPPRRVLETVVDAACMRAPEPLAIAYSPPFPAGDEGLYSE LAWRDRVAVVNESLVIYGALETDSGLYTLSVVGLSDEARQVASVVLVVEPAPVPTP TPDDYDEEDDAGVTNARRSAFPPQPPPRRPPVAPPTHPRVIPEVSHVRGVTVHMETL EAILFAPGETFGTNVSIHAIAHDDGPYAMDVVWMRFDVPSSCADMRIYEACLYHPQ LPECLSPADAPCAVSSWAYRLAVRSYAGCSRTTPPPRCFAEARMEPVPGLAWLAST VNLEFQHASPQHAGLYLCVYVDDHIHAWGHMTISTAAQYRNAVVEQHLPQRQ
  • an HSV-2 gE comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 18. In some embodiments, an HSV-2 gE has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 18.
  • an HSV-2 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: ABU45436.1, ABU45437.1, ABU45438.1, ABU45439.1, ABW83306.1, ABW83308.1, ABW83310.1, ABW83312.1, ABW83314.1, ABW83316.1, ABW83318.1, ABW83320.1, ABW83322.1, ABW83324.1, ABW83326.1, ABW83328.1, ABW83330.1, ABW83332.1, ABW83334.1, ABW83336.1, ABW83338.1, 66 11979815v1 P-627574-PC ABW83340.1, ABW83342.1, ABW83344.1, ABW83346.1, ABW83348.1, ABW83350.1, ABW83
  • an HSV gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises an IgG Fc-binding domain of the gE protein.
  • the gE domain encoded by RNA utilized in the methods and compositions of the present disclosure is any other gE domain known in the art to mediate binding to IgG Fc.
  • an HSV gE protein encoded by RNA utilized in the methods and compositions of the present disclosure comprises a gE domain involved in cell-to-cell spread.
  • an HSV gE fragment encoded by RNA fragment utilized in the methods and compositions of the present disclosure comprises an immune evasion domain.
  • the gE fragment encoded by RNA fragment utilized in the methods and compositions of the present disclosure comprises a portion of an immune evasion domain.
  • Each RNA encoding HSV-1 gE or HSV-2 gE protein or fragment thereof represents a separate embodiment of the present disclosure.
  • an HSV gE protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure is an immunogenic fragment.
  • a gE immunoprotective antigen need not be the entire protein.
  • the protective immune response generally involves, in another embodiment, an antibody response.
  • mutants, sequence conservative variants, and functional conservative variants of gE are useful in methods and compositions of the present disclosure, provided that all such variants retain the required immuno-protective effect.
  • the immunogenic fragment can comprise an immuno-protective gE antigen from any strain of HSV.
  • the immunogenic fragment can comprise sequence variants of HSV, as found in infected individuals.
  • an RNA of the present disclosure encodes an HSV polypeptide, or fragment thereof, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 1.
  • methods of the present disclosure comprise administering to a subject an HSV polypeptide, or fragment thereof, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 1.
  • Table 1 Exemplary amino acid sequences of HSV immunogens Sequence Amino acid sequence SEQ ID NO: 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name 69 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name 70 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name
  • methods of the present disclosure comprise administering to a subject an RNA comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 2.
  • Table 2 Exemplary nucleic acid sequences of HSV immunogens HSV-1 gD AAGUACGCCCUGGCCGACGCCUCCCUGAAGAUGGC 22 11979815v1 P-627574-PC GCCCCCGCCUUCGAGACCGCCGGCACCUACCUGCGC CUGGUGAAGAUCAACGACUGGACCGAGAUCACCCA 11979815v1 P-627574-PC CCGAGAACCAGCGCACCGUGGCCCUGUACUCCCUG AAGAUCGCCGGCUGGCACGGCCCCAAGCCCCCCUAC 11979815v1 P-627574-PC UAACUGGCACAUCCCCAGCAUCCAGGAUGUGGCCC CUCAUCAUUGA 11979815v1 P-627574-PC Version CGCGUCUACCACAUACAGCCUAGUCUUGAGGACCC 2.1 UUUUCAGCCACCGUCUAUCCCCAUUACCGUGUACU 11979815v1 P-627574-PC ACCUCACAAUAGCGUGGUAUCGAAUGGGC
  • an HSV glycoprotein, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure is an isoform of the sequence provided herein.
  • an HSV glycoprotein, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure is a variant of the sequence provided herein.
  • an HSV glycoprotein, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure is a fragment of the sequence provided herein.
  • a glycoprotein fragment encoded by RNA of the methods and compositions of the present disclosure comprises the ectodomain of the glycoprotein.
  • a glycoprotein fragment encoded by RNA of the methods and compositions of the present disclosure consists of the ectodomain of the glycoprotein.
  • a glycoprotein fragment encoded by RNA of the methods and compositions of the present disclosure comprises a fragment of the ectodomain of the glycoprotein.
  • the glycoprotein fragment may be any glycoprotein fragment known in the art.
  • a glycoprotein or immunogenic fragment encoded by an RNA utilized in the methods and compositions of the present disclosure may be from any strain of HSV.
  • an immunogenic fragment encoded by RNA utilized in the methods and compositions of the present disclosure may comprise sequence variants of HSV, as found in infected individuals.
  • variant refers to an amino acid or nucleic acid sequence (or in other embodiments, an organism or tissue) that is different from the majority of the population but is still sufficiently similar to the common mode to be considered to be one of them, for example splice variants.
  • the variant may a sequence conservative variant, while in another embodiment, the variant may be a functional conservative variant.
  • a variant may comprise an addition, deletion or substitution of one or more amino acids.
  • Immunune evasion domain refers, in some embodiments, to a domain that interferes with or reduces in vivo anti-HSV efficacy of anti-HSV antibodies (e.g., anti-gD antibodies).
  • the domain interferes or reduces in vivo anti-HSV efficacy of an anti-HSV immune response.
  • the domain reduces the immunogenicity of an HSV protein (e.g. gD) during subsequent infection.
  • the domain reduces the immunogenicity of an HSV protein during subsequent challenge.
  • the domain reduces the immunogenicity of HSV during subsequent challenge.
  • the domain reduces the immunogenicity of an HSV protein in the context of ongoing HSV infection.
  • the domain reduces the immunogenicity of HSV in the context of ongoing HSV infection.
  • the domain functions as an IgG Fc receptor.
  • the domain promotes antibody bipolar bridging, which in some embodiments, is a term that refers to an antibody molecule binding by its Fab domain to an HSV antigen and by its Fc domain to a separate HSV antigen, such as in some embodiments, gE, thereby blocking the ability of the Fc domain to activate complement.
  • the present disclosure also provides for modified RNA encoding analogs of HSV proteins or polypeptides, or fragments thereof. Analogs may differ from naturally occurring proteins or peptides by conservative amino acid sequence substitutions or by modifications which do not affect sequence, or by both.
  • an HSV glycoprotein, or an immunogenic fragment thereof, encoded by modified RNA of the present disclosure is homologous to a sequence set forth hereinabove, either expressly or by reference to a GenBank entry.
  • the terms “homology,” “homologous,” etc., when in reference to any protein or peptide, refer, in some embodiments, to a percentage of amino acid residues in the candidate sequence that are identical with the residues of a corresponding native polypeptide, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent homology, and not considering any conservative substitutions as part of the sequence identity. Methods and computer programs for the alignment are well known in the art.
  • “homology” refers to identity of a protein sequence encoded by an RNA to a sequence disclosed herein of greater than 70%. In another embodiment, the identity is greater than 72%. In another embodiment, the identity is greater than 75%. In another embodiment, the identity is greater than 78%. In another embodiment, the identity is greater than 80%. In another embodiment, the identity is greater than 82%. In another embodiment, the identity is greater than 83%. In another embodiment, the identity is greater than 85%. In another embodiment, the identity is greater than 87%. In another embodiment, the identity is greater than 128 11979815v1 P-627574-PC 88%. In another embodiment, the identity is greater than 90%. In another embodiment, the identity is greater than 92%.
  • isoform refers to a version of a molecule, for example, a protein, with only slight differences to another isoform of the same protein.
  • isoforms may be produced from different but related genes, or in another embodiment, may arise from the same gene by alternative splicing. In another embodiment, isoforms are caused by single nucleotide polymorphisms.
  • the RNA encoding a glycoprotein or glycoprotein fragment as described herein further encodes an antigenic tag.
  • the tag is a histidine (“His”) tag.
  • the His tag comprises 5 histidine residues.
  • the His tag comprises 6 histidine residues.
  • methods and compositions of the present disclosure utilize a chimeric molecule, comprising a fusion of an RNA encoding an HSV protein, or an immunogenic fragment thereof, with an RNA encoding a tag polypeptide that provides an epitope to which an anti-tag antibody can selectively bind.
  • the epitope tag is placed, in other embodiments, at the amino- or carboxyl-terminus of the protein or in an internal location therein.
  • the presence of such epitope-tagged forms of the recombinant HSV protein, or an immunogenic fragment thereof, is detected, in another embodiment, using an antibody against the tag polypeptide.
  • inclusion of the epitope tag enables the recombinant HSV protein, or an immunogenic fragment thereof, to be readily purified by affinity purification using an anti-tag antibody or another type of affinity matrix that binds to the epitope tag.
  • Various tag polypeptides and their respective antibodies are known in the art.
  • compositions of the present disclosure comprise an adjuvant, while in another embodiment, the compositions do not comprise an adjuvant.
  • adjuvant refers, in another embodiment, to compounds that, when administered to an individual or tested in vitro, increase the immune response to an antigen in the individual or test system to which the antigen is administered.
  • an immune adjuvant enhances an immune response to an antigen that is weakly immunogenic when administered alone, i.e., inducing no or weak antibody 129 11979815v1 P-627574-PC titers or cell-mediated immune response.
  • the adjuvant increases antibody titers to the antigen.
  • an RNA encoding a glycoprotein as described herein comprises a signal sequence encoding a signal peptide, e.g., that is functional in mammalian cells.
  • a signal sequence encodes a modified signal peptide (e.g., comprising amino acid substitutions or amino acid additions).
  • a signal sequence is a codon optimized signal sequence.
  • a utilized signal sequence is a heterologous signal sequence.
  • a heterologous signal sequence comprises or consists of a non-human signal sequence.
  • a heterologous signal sequence comprises or consists of a viral signal sequence.
  • a viral signal sequence comprises or consists of an HSV signal sequence (e.g., an HSV-1 or HSV-2 signal sequence).
  • a signal sequence comprises or consists of an HSV-1 signal sequence.
  • a signal sequence comprises or consists of an HSV-2 signal sequence.
  • a signal sequence is characterized by a length of about 15 to 30 amino acids.
  • a signal sequence encodes a signal peptide that preferably allows transport of an HSV-1 glycoprotein, an HSV-2 glycoprotein, an HSV-1 and HSV-2 glycoprotein, or an immunogenic fragment of an HSV-1 and/or HSV-2 glycoprotein, with which it is associated into a defined cellular compartment, preferably a cell surface, endoplasmic reticulum (ER) or endosomal- lysosomal compartment.
  • a signal sequence is the native signal sequence of the encoded glycoprotein.
  • a signal sequence is or comprises an HSV glycoprotein D (gD) signal sequence (e.g., an HSV-1 or HSV-2 gD signal sequence).
  • gD HSV glycoprotein D
  • a signal peptide encoded by a signal sequence is or comprises an HSV-2 gD signal peptide (SEQ ID NO: 29).
  • an HSV-2 gD signal peptide comprises KY (SEQ ID NO: 30), KYA (SEQ ID NO: 31, KYAL (SEQ ID NO: 32), or KYALA (SEQ ID NO: 33) at the C terminus of the signal peptide.
  • a signal peptide is or comprises an HSV-1 gD signal 130 11979815v1 P-627574-PC peptide (SEQ ID NO: 34).
  • an HSV-1 gD signal peptide comprises KY at the C terminus of the signal sequence (SEQ ID NO: 35).
  • a signal sequence is or comprises an HSV glycoprotein C (gC) signal sequence (e.g., an HSV-1 or HSV-2 gC signal sequence).
  • a signal peptide encoded by a signal sequence is or comprises an HSV-2 gC signal peptide (SEQ ID NO: 36).
  • a signal peptide is or comprises an HSV-1 gC signal peptide (SEQ ID NO: 37).
  • a signal sequence is or comprises an HSV glycoprotein E (gE) signal sequence (e.g., an HSV-1 or HSV-2 gE signal sequence).
  • a signal peptide encoded by a signal sequence is or comprises an HSV-2 gE signal sequence (SEQ ID NO: 38).
  • a signal peptide is or comprises an HSV-1 gE signal peptide (SEQ ID NO: 39).
  • an HSV-2 gE signal peptide comprises RTS at the C terminus of the signal peptide (SEQ ID NO: 40).
  • an HSV-2 gE signal peptide comprises A20V, A21V, and A22V substitutions (SEQ ID NO: 41).
  • a signal sequence is or comprises an HSV gB signal sequence (e.g., an HSV-1 or HSV-2 gB signal sequence).
  • a signal peptide encoded by a signal sequence is or comprises an HSV-1 gB signal peptide (SEQ ID NO: 42). In some embodiments, a signal peptide encoded by a signal sequence is or comprises an HSV-2 gB signal peptide (SEQ ID NO: 43). In some embodiments, an HSV-1 gB signal peptide comprises AP at the C terminus of the signal peptide (SEQ ID NO: 44). [0254] In some embodiments, a signal sequence is or comprises an HSV gI signal sequence (e.g., an HSV-1 or HSV-2 gI signal sequence).
  • a signal peptide encoded by a signal sequence is or comprises an HSV-1 gI signal peptide (SEQ ID NO: 45).
  • an HSV-2 gI signal sequence is or comprises an HSV-2 gI signal peptide (SEQ ID NO: 249).
  • a signal peptide encoded by a signal sequence is or comprises amino acid residues 1-18 of the wild-type sequence (SEQ ID NO: 46).
  • an HSV-2 gI signal peptide comprises an additional leucine residue at the C terminus of the signal peptide (SEQ ID NO: 47).
  • a signal sequence is a heterologous signal sequence.
  • a heterologous signal peptide encoded by a heterologous signal sequence comprises an IL-2 signal peptide (SEQ ID NO: 220). In some embodiments, a heterologous signal peptide encoded by a heterologous signal sequence comprises or consists of an azurocidin signal peptide 131 11979815v1 P-627574-PC (SEQ ID NO: 221). In some embodiments, a heterologous signal peptide encoded by a heterologous signal sequence comprises or consists of an MHC Class II signal peptide (SEQ ID NO: 222). In some embodiments, a signal sequence comprises or consists of an Ebola virus signal sequence.
  • an Ebola virus signal peptide comprises or consists of an Ebola virus spike glycoprotein (SGP) signal peptide (SEQ ID NO: 48).
  • SGP Ebola virus spike glycoprotein
  • an RNA encoding a glycoprotein as described herein does not comprise a signal sequence.
  • an RNA as described herein comprises only an ectodomain.
  • a signal sequence encodes an exemplary signal peptide listed in Table 3, or a signal peptide having 1, 2, 3, 4, or 5 amino acid differences relative thereto.
  • a signal peptide is an exemplary signal peptide selected from those listed in Table 3 and functionally connected to the N-terminus of an exemplary HSV immunogen selected from those listed in Table 1.
  • Table 3 Exemplary signal peptides Sequence Name Amino Acid Sequence SEQ ID NO: 11979815v1 P-627574-PC Sequence Name Amino Acid Sequence SEQ ID NO: HSV-2 gE Signal Sequence MARGAGLVFFVGVWVVSCLVVVP 41 [ ed in T able 4, or a signal sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence listed in Table 4.
  • an exemplary signal sequence is selected from those listed in Table 4 and functionally connected (i.e., in frame) to the 5' end of an exemplary HSV immunogen nucleic acid sequence selected from those listed in Table 2.
  • a nucleotide sequence encodes a protein, wherein the protein comprises an HSV-2 glycoprotein antigen, or immunogenic fragment thereof, and a signal peptide.
  • the protein comprises an HSV-2 glycoprotein antigen, or immunogenic fragment thereof, and a signal peptide.
  • a protein of the present disclosure comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 5. Table 5.
  • an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gC protein or immunogenic fragment thereof. In some embodiments, an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gD protein or immunogenic fragment thereof. In some embodiments, an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gE protein or immunogenic fragment thereof. [0261] Exemplary RNA comprising a signal sequence (i.e., encoding a signal peptide) and a sequence encoding an HSV-2 glycoprotein are shown in Table 6 below.
  • an RNA of the present disclosure comprises a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6.
  • an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gD protein or immunogenic fragment thereof and a signal sequence. In some embodiments, an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gE protein or immunogenic fragment thereof and a signal sequence. [0263] In some embodiments, nucleotide sequences described herein can comprise a nucleotide sequence that encodes a 5’UTR and/or a 3’ UTR. In some embodiments, polynucleotides described herein can comprise a nucleotide sequence that encodes a polyA tail.
  • nucleotide sequences described herein may comprise a 5’ cap, which may be incorporated during transcription, or joined to a nucleotide sequence post-transcription.
  • a nucleotide sequence provided herein encodes one or more glycoproteins (e.g., glycoprotein C (gC), glycoprotein D (gD), glycoprotein E (gE), or a combination thereof), or an immunogenic fragment thereof.
  • an RNA comprises a 5’ cap, a 5’UTR, a nucleotide sequence that encodes one or more glycoproteins (e.g., glycoprotein C (gC), glycoprotein D (gD), glycoprotein E (gE), or a combination thereof), or an immunogenic fragment thereof, a 3’ UTR, and a polyA tail.
  • mRNA messenger RNA
  • a structural feature of messenger RNA (mRNA) is a cap structure at the five-prime end (5').
  • Natural eukaryotic mRNA comprise a 7-methylguanosine cap linked to the mRNA via a 5 ⁇ to 5 ⁇ -triphosphate bridge resulting in a cap0 structure (m7GpppN).
  • RNA capping is well researched and is described, e.g., in Decroly E et al. (2012) Nature Reviews 10: 51-65; and in Ramanathan A. et al., (2016) Nucleic Acids Res; 44(16): 7511–7526, the entire contents of each of which are hereby incorporated by reference.
  • a 5’-cap structure which may be suitable in the context of the present invention is a cap0 (methylation of the first nucleobase, e.g., m7GpppN), cap1 (additional methylation of the ribose of the adjacent nucleotide of m7GpppN), cap2 (additional methylation of the ribose of the 2nd nucleotide downstream of the m7GpppN), cap3 (additional methylation of the ribose of the 3rd nucleotide downstream of the m7GpppN), cap4 (additional methylation of the ribose of the 4th nucleotide downstream of the m7GpppN), ARCA (“anti-reverse cap analogue”), modified ARCA (e.g.
  • RNA e.g., mRNA
  • 5'-cap refers to a structure found on the 5'-end of an RNA, e.g., mRNA, and generally includes a guanosine nucleotide connected to an RNA, e.g., mRNA, via a 5'- to 5'-triphosphate linkage (also referred to as Gppp or G(5')ppp(5')).
  • a guanosine nucleoside included in a 5’ cap may be modified, for example, by methylation at one or more positions (e.g., at the 7-position) on a base (guanine), and/or by methylation at one or more positions of a ribose.
  • a guanosine nucleoside included in a 5’ cap comprises a 3’O methylation at a ribose (3’OMeG).
  • a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine (m7G).
  • a guanosine nucleoside included in a 5' cap comprises methylation at the 7'-position of guanine and a 3' O methylation at a ribose (m7(3'OMeG)).
  • m7(3'OMeG) a 3' O methylation at a ribose
  • providing an RNA with a 5'-cap disclosed herein may be achieved by in vitro transcription, in which a 5'-cap is co-transcriptionally incorporated into an RNA strand, or may be attached to an RNA post-transcriptionally using capping enzymes.
  • co-transcriptional capping with a cap disclosed herein improves the capping efficiency of an RNA compared to co-transcriptional capping with an appropriate reference comparator.
  • improving capping efficiency can increase a translation efficiency and/or translation rate of an RNA, and/or increase expression of an encoded protein.
  • alterations to polynucleotides generate a non-hydrolyzable cap structure which can, for example, prevent decapping and increase RNA half-life.
  • a utilized 5' cap is a cap0, a cap1, or cap2 structure. See, e.g., Fig. 1 of Ramanathan A et al., and Fig. 1 of Decroly E et al., each of which is incorporated herein by reference in its entirety.
  • an RNA described herein comprises a cap1 structure.
  • an RNA described herein comprises a cap2 structure.
  • an RNA described herein comprises a cap0 structure.
  • a cap0 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G). In some embodiments, such a cap0 structure is connected to an RNA via a 5'- to 5'-triphosphate linkage and is also referred to herein as (m7)Gppp. In some embodiments, a cap0 structure comprises a guanosine nucleoside methylated at the 2'-position of the ribose of guanosine. In some embodiments, a cap0 structure comprises a guanosine nucleoside methylated at the 3'-position of the ribose of guanosine.
  • a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine and at the 2'-position of the ribose ((m27,2'-O)G). In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine and at the 2'-position of the ribose ((m27,3'-O)G).
  • a cap1 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and optionally methylated at the 2' or 3' position of the ribose, and a 2'O methylated first nucleotide in an RNA ((m2'-O)N1).
  • a cap1 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and the 3' position of the ribose, and a 2'O methylated first nucleotide in an RNA ((m2'-O)N1).
  • a cap1 structure is connected to an RNA via a 5'- to 5'-triphosphate linkage and is 246 11979815v1 P-627574-PC also referred to herein as, e.g., ((m7)Gppp(2'-O)N1) or (m27,3’-O)Gppp(2'-O)N1), wherein N1 is as defined and described herein.
  • a cap1 structure comprises a second nucleotide, N2, which is at position 2 and is chosen from A, G, C, or U, e.g., (m7)Gppp(2'- O)N1pN2 or (m27,3’-O)Gppp(2'-O)N1pN2 , wherein each of N1 and N2 is as defined and described herein.
  • a cap2 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and optionally methylated at the 2' or 3' position of the ribose, and 2'O methylated first and second nucleotides in an RNA ((m2’-O)N1p(m2’-O)N2).
  • a cap2 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and the 3' position of the ribose, and 2'O methylated first and second nucleotides in an RNA.
  • a cap2 structure is connected to an RNA via a 5'- to 5'- triphosphate linkage and is also referred to herein as, e.g., ((m7)Gppp(2'-O)N1p(2'-O)N2) or (m27,3’-O)Gppp(2'-O)N1p(2'-O)N2), wherein each of N1 and N2 is as defined and described herein.
  • the 5' cap is a dinucleotide cap structure.
  • the 5' cap is a dinucleotide cap structure comprising N1, wherein N1 is as defined and described herein.
  • the 5' cap is a dinucleotide cap G*N1, wherein N1 is as defined above and herein, and G* comprises a structure of formula (I): wherein each R 2 and R 3 is -OH or -OCH 3 ; and X is O or S.
  • R 2 is -OH.
  • R 2 is -OCH 3 .
  • R3 is -OH.
  • R3 is -OCH3.
  • R2 is -OH and R3 is -OH.
  • R2 is -OH and R3 is -CH3.
  • R2 is - CH3 and R3 is -OH.
  • R2 is -CH3 and R3 is -CH3.
  • X is O.
  • X is S. 247 11979815v1 P-627574-PC
  • the 5' cap is a dinucleotide cap0 structure (e.g., (m7)GpppN1, (m27,2’-O)GpppN1, (m27,3’-O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’- O)GppSpN1), wherein N1 is as defined and described herein.
  • the 5' cap is a dinucleotide cap0 structure (e.g., (m7)GpppN1, (m27,2’-O)GpppN1, (m27,3’-O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’-O)GppSpN1), wherein N1 is G.
  • a dinucleotide cap0 structure e.g., (m7)GpppN1, (m27,2’-O)GpppN1, (m27,3’-O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’-O)GppSpN1
  • N1 is G.
  • the 5' cap is a dinucleotide cap0 structure (e.g., (m7)GpppN1, (m27,2’-O)GpppN1, (m27,3’-O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’-O)GppSpN1), wherein N1 is A, U, or C.
  • N1 is A, U, or C.
  • the 5' cap is a dinucleotide cap1 structure (e.g., (m7)Gppp(m2’-O)N1, (m27,2’-O)Gppp(m2’-O)N1, (m27,3’-O)Gppp(m2’-O)N1, (m7)GppSp(m2’-O)N1, (m27,2’-O)GppSp(m2’-O)N1, or (m27,3’-O)GppSp(m2’-O)N1), wherein N 1 is as defined and described herein.
  • N 1 is as defined and described herein.
  • the 5' cap is selected from the group consisting of (m 7 )GpppG (“Ecap0”), (m 7 )Gppp(m 2’-O )G (“Ecap1”), (m 2 7,3’-O )GpppG (“ARCA” or “D1”), and (m 2 7,2’-O )GppSpG (“beta-S-ARCA”).
  • the 5' cap is (m 7 )GpppG (“Ecap0”), having a structure of formula (II): [0277]
  • the 5' cap is (m 7 )Gppp(m 2’-O )G (“Ecap1”), having a structure of formula (III): P-627574-PC [0278]
  • the 5' cap is (m 2 7,3’-O )GpppG (“ARCA” or “D1”), having a structure of formula (IV): OH O O some cap ARCA”), having a structure of formula (V): O OH O [0280]
  • the 5' cap is a trinucleotide cap structure.
  • the 5' cap is a trinucleotide cap structure comprising N 1 pN 2 , wherein N 1 and N 2 are as defined and described herein.
  • the 5' cap is a dinucleotide cap G*N1pN2, wherein N1 and N 2 are as defined above and herein, and G* comprises a structure of formula (VI): defined and described herein.
  • the 5' cap is a trinucleotide cap0 structure (e.g., (m 7 )GpppN 1 pN 2 , (m 2 7,2’-O )GpppN 1 pN 2 , or (m 2 7,3’-O )GpppN 1 pN 2 ), wherein N 1 and N 2 are as defined and described herein).
  • the 5' cap is a trinucleotide cap1 structure (e.g., (m 7 )Gppp(m 2’- O )N 1 pN 2 , (m 2 7,2’-O )Gppp(m 2’-O )N 1 pN 2 , (m 2 7,3’-O )Gppp(m 2’-O )N 1 pN 2 ), wherein N 1 and N 2 are as defined and described herein.
  • the 5' cap is a trinucleotide cap2 structure (e.g., (m 7 )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 , (m 2 7,2’-O )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 , (m 2 7,3’-O )Gppp(m 2’- O )N 1 p(m 2’-O )N 2 ), wherein N 1 and N 2 are as defined and described herein.
  • the 5' cap is selected from the group consisting of (m2 7,3’-O )Gppp(m 2’-O )ApG (“CleanCap AG”, “CC413”), (m 2 7,3’-O )Gppp(m 2’-O )GpG (“CleanCap GG”), (m 7 )Gppp(m 2’-O )ApG, (m 7 )Gppp(m 2’- O )GpG, (m 2 7,3’-O )Gppp(m 2 6,2’-O )ApG, and (m 7 )Gppp(m 2’-O )ApU.
  • the 5' cap is (m 2 7,3’-O )Gppp(m 2’-O )ApG (“CleanCap AG”, “CC413”), having a structure of formula (VII): [0283] In some embodiments, the 5' cap is (m 2 7,3’-O )Gppp(m 2’-O )GpG (“CleanCap GG”), having a structure of formula (VIII): 250 11979815v1 P-627574-PC some cap a structure of formula (IX): OH OH NH 2 [0285] In some embodiments, the 5' cap is (m 7 )Gppp(m 2’-O )GpG, having a structure of formula (X): 251 11979815v1 P-627574-PC or a [0286] In some embodiments, the 5' cap is (m2 7,3’-O )Gppp(m2 6,2’-O )ApG, having
  • the 5' cap is a tetranucleotide cap structure comprising N 1 pN 2 pN 3 , wherein N 1 , N 2 , and N 3 are as defined and described herein.
  • the 5' cap is a tetranucleotide cap G*N 1 pN 2 pN 3 , wherein N 1 , N 2 , and N 3 are as defined above and herein, and G* comprises a structure of formula (XIII): or a salt thereof, wherein R 2 , R 3 , and X are as defined and described herein.
  • the 5' cap is a tetranucleotide cap0 structure (e.g.
  • the 5’ cap is a tetranucleotide Cap1 structure (e.g., (m 7 )Gppp(m 2’-O )N1pN2pN3, (m2 7,2’-O )Gppp(m 2’-O )N1pN2pN3, (m2 7,3’-O )Gppp(m 2’- O )N 1 pN 2 N 3 ), wherein N 1 , N 2 , and N 3 are as defined and described herein.
  • tetranucleotide Cap1 structure e.g., (m 7 )Gppp(m 2’-O )N1pN2pN3, (m2 7,2’-O )Gppp(m 2’-O )N1pN2pN3, (m2 7,3’-O )Gppp(m 2’- O )N 1 pN 2 N 3
  • N 1 , N 2 , and N 3 are as defined and described herein.
  • the 5' cap is a tetranucleotide Cap2 structure (e.g., (m 7 )Gppp(m 2’-O )N1p(m 2’-O )N2pN3, (m2 7,2’- O )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 pN 3 , (m 2 7,3’-O )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 pN 3 ), wherein N 1 , N 2 , and N 3 are as defined and described herein.
  • N 1 , N 2 , and N 3 are as defined and described herein.
  • the 5' cap is selected from the group 253 11979815v1 P-627574-PC consisting of (m 2 7,3’-O )Gppp(m 2’-O )Ap(m 2’-O )GpG, (m 2 7,3’-O )Gppp(m 2’-O )Gp(m 2’-O )GpC, (m 7 )Gppp(m 2’-O )Ap(m 2’-O )UpA, and (m 7 )Gppp(m 2’-O )Ap(m 2’-O )GpG.
  • the 5' cap is (m 2 7,3’-O )Gppp(m 2’-O )Ap(m 2’-O )GpG, having a structure of formula (XIV): [0291] In some embodiments, the 5' cap is (m27 ,3’-O )Gppp(m 2’-O )Gp(m 2’-O )GpC, having a structure of formula (XV): 254 11979815v1 P-627574-PC some cap having a structure of formula (XVI): P-627574-PC [0293] In some embodiments, the 5' cap is (m 7 )Gppp(m 2’-O )Ap(m 2’-O )GpG, having a structure of formula (XVII): or a salt thereof.
  • a 5' UTR utilized in accordance with the present disclosure comprises a cap proximal sequence, e.g., as disclosed herein.
  • a cap proximal sequence comprises a sequence adjacent to a 5' cap.
  • a cap proximal sequence comprises nucleotides in positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.
  • a cap structure comprises one or more polynucleotides of a cap proximal sequence.
  • a cap structure comprises an m7 guanosine cap and nucleotide +1 (N1) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m7 guanosine cap and nucleotide +2 (N2) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m7 guanosine cap and nucleotides +1 and +2 (N1 and N2) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m7 guanosine cap and nucleotides +1, +2, and +3 (N1, N2, and N3) of an RNA polynucleotide.
  • one or more residues of a cap proximal sequence may be included in an RNA by virtue of having been included in a cap entity (e.g., a cap1 or cap2 structure, etc.); alternatively, in some embodiments, at least some of the residues in a cap proximal sequence may be enzymatically added (e.g., by a polymerase such as a T7 polymerase).
  • the 5'’ cap is a dinucleotide cap structure, wherein the cap proximal sequence comprises N1 of the 5’ cap, where N1 is any nucleotide, e.g., A, C, G or U.
  • the 5' cap is a trinucleotide cap structure (e.g., the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N1 and N2 of the 5' cap, wherein N1 and N2 are independently any nucleotide, e.g., A, C, G or U.
  • the 5' cap is a tetranucleotide cap structure (e.g., the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N1, N2, and N3 of the 5' cap, wherein N1, N2, and N3 are any nucleotide, e.g., A, C, G or U.
  • the cap proximal sequence comprises N1, N2, and N3 of the 5' cap, wherein N1, N2, and N3 are any nucleotide, e.g., A, C, G or U.
  • a cap proximal sequence comprises N1 of a the 5' cap, and N2, N3, N4 and N5, wherein N1 to N5 correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.
  • a cap proximal sequence comprises N1 and N2 of a the 5' cap, and N3, N4 and N5, wherein N1 to N5 correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.
  • a cap proximal sequence comprises N1, N2, and N3 of a the 5’ cap, and N4 and N5, wherein N1 to N5 correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.
  • N1 is A.
  • N1 is C.
  • N1 is G.
  • N1 is U.
  • N2 is A.
  • N2 is C.
  • N2 is G.
  • N2 is U.
  • N3 is A.
  • N3 is C. In some embodiments, N3 is G. In some embodiments, N3 is U. In some embodiments, N4 is A. In some embodiments, N4 is C. In some embodiments, N4 is G. In some embodiments, N4 is U. In some embodiments, N5 is A. In some embodiments, N5 is C. In some embodiments, N5 is G. In some embodiments, N5 is U. It will be understood that, each of the embodiments described above and herein (e.g., for N1 through N5) may be taken singly or in combination and/or may be combined with other embodiments of variables described above and herein (e.g., 5' caps).
  • a cap proximal sequence comprises A1 and G2 of the Cap1 structure, and a sequence comprising: A 3 A 4 U 5 at positions +3, +4 and +5 respectively of the nucleotide sequence. 3. 5’ UTR [0301]
  • an RNA utilized in accordance with the present disclosure comprises a 5'-UTR.
  • a 5’-UTR may comprise a plurality of distinct sequence elements; in some embodiments, such plurality may be or comprise multiple copies of one or more particular sequence elements (e.g., as may be from a particular source or otherwise known as a functional or characteristic sequence element).
  • a 5’ UTR comprises multiple different sequence elements.
  • the term “untranslated region” or “UTR” is commonly used in the art to refer to a region in a DNA molecule which is transcribed but is not translated into an amino acid sequence, or to the corresponding region in an RNA polynucleotide, such as an mRNA molecule.
  • An untranslated region (UTR) can be present 5' (upstream) of an open reading frame (5'-UTR) and/or 3' (downstream) of an open reading frame (3'-UTR).
  • the terms “five prime untranslated region” or “5' UTR” refer to a sequence of a nucleotide sequence between the 5' end of the nucleotide sequence (e.g., a transcription start site) and a start codon of a coding region of the nucleotide sequence.
  • “5' UTR” refers to a sequence of a nucleotide sequence that begins at the 5' end of the nucleotide sequence (e.g., a transcription start site) and ends one nucleotide (nt) before a start codon (usually AUG) of a coding region of the nucleotide sequence, e.g., in its natural context.
  • a 5' UTR comprises a Kozak sequence.
  • a 5'-UTR is downstream of the 5'-cap (if present), e.g., directly adjacent to the 5'-cap.
  • a 5' UTR disclosed herein comprises a cap proximal sequence, e.g., as defined and described herein.
  • a cap proximal sequence comprises a sequence adjacent to a 5' cap.
  • Exemplary 5' UTRs include a human alpha globin (hAg) 5'UTR or a fragment thereof, a TEV 5' UTR or a fragment thereof, a HSP705' UTR or a fragment thereof, or a c-Jun 5' UTR or a fragment thereof.
  • an RNA disclosed herein comprises a hAg 5' UTR or a fragment thereof.
  • an RNA disclosed herein comprises a 5' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% 258 11979815v1 P-627574-PC identity to a 5' UTR with the sequence AGAATAAACTAGTATTCTTCTGGTCCCCACAGACTCAGAGAGAACCCGCCACC (SEQ ID NO: 210).
  • an RNA disclosed herein comprises a 5' UTR having the sequence as set forth in SEQ ID NO: 210.
  • an RNA disclosed herein comprises a 5' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 5’ UTR with the sequence AACUAGUAUUCUUCUGGUCCCCACAGACUCAGAGAGAACCCGCCACC (SEQ ID NO: 211) (hAg-Kozak/5'UTR).
  • an RNA disclosed herein comprises a 5' UTR having the sequence as set forth in SEQ ID NO 211. 4.
  • a polynucleotide e.g., DNA, RNA
  • a polyA sequence is situated downstream of a 3'-UTR, e.g., adjacent to a 3'-UTR.
  • poly(A) sequence or “poly-A tail” refers to an uninterrupted or interrupted sequence of adenylate residues which is typically located at the 3'-end of an RNA polynucleotide.
  • Poly(A) sequences are known to those of skill in the art and may follow the 3’- UTR in the RNAs described herein.
  • An uninterrupted poly(A) sequence is characterized by consecutive adenylate residues. In nature, an uninterrupted poly(A) sequence is typical.
  • polynucleotides disclosed herein comprise an uninterrupted poly(A) sequence.
  • polynucleotides disclosed herein comprise interrupted poly(A) sequence.
  • RNAs disclosed herein can have a poly(A) sequence attached to the free 3'- end of the RNA by a template-independent RNA polymerase after transcription or a poly(A) sequence encoded by DNA and transcribed by a template-dependent RNA polymerase.
  • a poly(A) sequence of about 120 A nucleotides has a beneficial influence on the levels of RNA in transfected eukaryotic cells, as well as on the levels of protein that are translated from an open reading frame that is present upstream (5') of the poly(A) sequence (Holtkamp et al., 2006, Blood, vol.108, pp.4009-4017, which is herein incorporated by reference).
  • a poly(A) sequence in accordance with the present disclosure is not limited to a particular length; in some embodiments, a poly(A) sequence is any length.
  • a poly(A) sequence comprises, essentially consists of, or consists of at least 20, at 259 11979815v1 P-627574-PC least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 A nucleotides, and, in particular, about 120 A nucleotides.
  • nucleotides in the poly(A) sequence typically at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by number of nucleotides in the poly(A) sequence are A nucleotides, but permits that remaining nucleotides are nucleotides other than A nucleotides, such as U nucleotides (uridylate), G nucleotides (guanylate), or C nucleotides (cytidylate).
  • a poly(A) sequence is attached during RNA transcription, e.g., during preparation of in vitro transcribed RNA, based on a DNA template comprising repeated dT nucleotides (deoxythymidylate) in the strand complementary to the coding strand.
  • the DNA sequence encoding a poly(A) sequence (coding strand) is referred to as a poly(A) cassette.
  • the poly(A) cassette present in the coding strand of DNA essentially consists of dA nucleotides, but is interrupted by a random sequence of the four nucleotides (dA, dC, dG, and dT). Such a random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides in length.
  • a cassette is disclosed in WO 2016/005324 A1, hereby incorporated by reference. Any poly(A) cassette disclosed in WO 2016/005324 A1, which is incorporated herein by reference in its entirety, may be used in accordance with the present disclosure.
  • a poly(A) cassette that essentially consists of dA nucleotides, but is interrupted by a random sequence having an equal distribution of the four nucleotides (dA, dC, dG, dT) and having a length of e.g., 5 to 50 nucleotides shows, at the DNA level, constant propagation of plasmid DNA in E. coli and is still associated, at the RNA level, with the beneficial properties with respect to supporting RNA stability and translational efficiency is encompassed.
  • the poly(A) sequence contained in an RNA polynucleotide described herein essentially consists of A nucleotides, but is interrupted by a random sequence of the four nucleotides (A, C, G, U).
  • Such a random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides in length.
  • no nucleotides other than A nucleotides flank a poly(A) sequence at its 3'-end, i.e., the poly(A) sequence is not masked or followed at its 3'-end by a nucleotide other than A.
  • the poly(A) sequence may comprise at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 260 11979815v1 P-627574-PC nucleotides.
  • the poly(A) sequence may essentially consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence may consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence comprises at least 100 nucleotides. In some embodiments, the poly(A) sequence comprises about 150 nucleotides. In some embodiments, the poly(A) sequence comprises about 120 nucleotides.
  • a poly(A) sequence comprises a specific number of adenosines, such as about 50 or more, about 60 or more, about 70 or more, about 80 or more, about 90 or more, about 100 or more, about 120, or about 150 or about 200.
  • a poly(A) sequence of an RNA may comprise 200 A residues or less.
  • a poly(A) sequence of an RNA may comprise about 200 A residues.
  • a poly(A) sequence of an RNA may comprise 180 A residues or less.
  • a poly(A) sequence of an RNA may comprise about 180 A residues.
  • a poly(A) sequence may comprise 150 residues or less.
  • RNA comprises a poly(A) sequence comprising the nucleotide sequence of: AAAAAAAAAAAAAAAAAAAAAAAAAAGCATATGACTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
  • a linker comprises the nucleotide sequence GCATATGAC.
  • an RNA comprises a poly(A) sequence comprising the nucleotide sequence of: AAAAAAAAAAAAAAAAAAAAAAAAAAAAGCAUAUGACUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
  • a poly(A) sequence comprises a plurality of A residues interrupted by a linker.
  • a linker comprises the nucleotide sequence GCAUAUGAC. 261 11979815v1 P-627574-PC 5.
  • 3' UTR [0318]
  • an RNA utilized in accordance with the present disclosure comprises a 3'-UTR.
  • the terms “three prime untranslated region,” “3' untranslated region,” or “3' UTR” refer to a sequence of an mRNA molecule that begins following a stop codon of a coding region of an open reading frame sequence.
  • the 3' UTR begins immediately after a stop codon of a coding region of an open reading frame sequence, e.g., in its natural context. In other embodiments, the 3' UTR does not begin immediately after stop codon of the coding region of an open reading frame sequence, e.g., in its natural context.
  • the term “3'- UTR” preferably does not include the poly(A) sequence.
  • the 3'-UTR is upstream of the poly(A) sequence (if present), e.g. directly adjacent to the poly(A) sequence.
  • an RNA disclosed herein comprises a 3'UTR comprising an F element and/or an I element.
  • a 3' UTR or a proximal sequence thereto comprises a restriction site.
  • a restriction site is a BamHI site.
  • a restriction site is an XhoI site.
  • an RNA construct comprises an F element.
  • an F element sequence is a 3' UTR of amino-terminal enhancer of split (AES).
  • an RNA disclosed herein comprises a 3' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 3’ UTR with the sequence of CTGGTACTGCATGCACGCAATGCTAGCTGCCCCTTTCCCGTCCTGGGTACCCCGAGT CTCCCCCGACCTCGGGTCCCAGGTATGCTCCCACCTCCACCTGCCCCACTCACCACC TCTGCTAGTTCCAGACACCTCCCAAGCACGCAGCAATGCAGCTCAAAACGCTTAGCC TAGCCACACCCACGGGAAACAGCAGTGATTAACCTTTAGCAATAAACGAAAGTT TAACTAAGCTATACTAACCCCAGGGTTGGTCAATTTCGTGCCAGCCACACC (SEQ ID NO: 216).
  • an RNA disclosed herein comprises a 3' UTR with the sequence of SEQ ID NO: 216. [0322] In some embodiments, an RNA disclosed herein comprises a 3' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 3’ UTR with the sequence of CUGGUACUGCAUGCACGCAAUGCUAGCUGCCCCUUUCCCGUCCUGGGUACCCCGA GUCUCCCCCGACCUCGGGUCCCAGGUAUGCUCCCACCUCCACCUGCCCCACUCACC ACCUCUGCUAGUUCCAGACACCUCCCAAGCACGCAGCAAUGCAGCUCAAAACGCU 262 11979815v1 P-627574-PC UAGCCUAGCCACACCCCCACGGGAAACAGCAGUGAUUAACCUUUAGCAAUAAACG AAAGUUUAACUAAGCUAUACUAACCCCAGGGUUGGUCAAUUUCGU
  • an RNA disclosed herein comprises a 3' UTR with the sequence of SEQ ID NO: 217.
  • a 3' UTR is an FI element as described in WO2017/060314, which is herein incorporated by reference in its entirety.
  • the present disclosure provides compositions comprising modified RNAs and methods of use thereof.
  • the modified RNA comprises one or more modified nucleoside residues.
  • an RNA comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 2, Table 4, or Table 6, comprises one or more modified nucleoside residues.
  • an RNA as described herein refers to a messenger RNA.
  • all uridine residues are modified as described herein.
  • one or more of the RNAs as described herein are nucleoside modified RNAs.
  • the modified nucleoside of the methods and compositions of the present disclosure is m5C (5-methylcytidine).
  • the modified nucleoside is m5U (5-methyluridine).
  • the modified nucleoside is m6A (N6- methyladenosine).
  • the modified nucleoside is s2U (2-thiouridine).
  • the modified nucleoside is ⁇ (pseudouridine).
  • the modified nucleoside is Um (2'-O-methyluridine).
  • a modified nucleoside is m 1 A (1-methyladenosine), m 2 A (2- methyladenosine), m 6 A (N6-methyladenosine), Am (2'-O-methyladenosine), ms 2 m 6 A (2- methylthio-N6-methyladenosine), i 6 A (N6-isopentenyladenosine), ms 2 i 6 A (2-methylthio-N6- isopentenyladenosine), io 6 A (N6-(cis-hydroxyisopentenyl)adenosine), ms 2 io 6 A (2-methylthio- N6-(cis-hydroxyisopentenyl) adenosine), g 6 A (N6-glycinylcarbamoyladenosine), t 6 A (N6-N6-glycinylcar
  • modified nucleoside residues are pseudouridine or pseudouridine family residues.
  • the modified RNA comprises pseudouridine residues.
  • pseudouridine refers to the C-glycoside isomer of the nucleoside uridine.
  • pseudouridine residues comprise m 1 acp 3 ⁇ (1-methyl-3-(3-amino-5- carboxypropyl)pseudouridine, m 1 ⁇ (1-methylpseudouridine), ⁇ m (2′-O-methylpseudouridine, m 5 D (5-methyldihydrouridine), m 3 ⁇ (3-methylpseudouridine), or a combination thereof.
  • said pseudouridine residues comprise 1-methylpseudouridine residues instead of uridine.
  • modified nucleoside residues are pseudouridine analogues.
  • a "pseudouridine analog” is any modification, variant, isoform or derivative of pseudouridine.
  • pseudouridine analogs include but are not limited to 1- carboxymethyl-pseudouridine, 1-propynyl-pseudouridine, 1-taurinomethyl-pseudouridine, 1- taurinomethyl-4-thio-pseudouridine, 1-methylpseudouridine (m 1 ⁇ ), 1-methyl-4-thio- pseudouridine (m 1 s 4 ⁇ ), 4-thio-1-methyl-pseudouridine, 3-methyl-pseudouridine (m 3 ⁇ ), 2-thio-1- methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydropseudouridine, 2-thio-dihydropseudouridine, 2-methoxyuridine,
  • a modified nucleobase is a modified uracil.
  • Exemplary nucleobases and nucleosides having a modified uracil include pseudouridine ( ⁇ ), pyridin-4-one ribonucleoside, 5-aza-uridine, 6-aza-uridine, 2-thio-5-aza-uridine, 2-thio-uridine (s 2 U), 4-thio- uridine (s 4 U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxy-uridine (ho 5 U), 5- 265 11979815v1 P-627574-PC aminoallyl-uridine, 5-halo-uridine (e.g., 5-iodo-uridine or 5-bromo-uridine), 3-methyl-uridine (m 3 U), 5-methoxy-uridine (mo 5 U), uridine 5-oxyacetic acid (cmo 5 U), uridine 5-oxyacetic acid methyl ester (mcmo
  • a modified nucleobase is a modified cytosine.
  • exemplary nucleobases and nucleosides having a modified cytosine include 5-aza-cytidine, 6-aza-cytidine, pseudoisocytidine, 3-methyl-cytidine (m 3 C), N4-acetyl-cytidine (ac 4 C), 5-formyl-cytidine (f 5 C), N4-methyl-cytidine (m 4 C), 5-methyl-cytidine (m 5 C), 5-halo-cytidine (e.g., 5-iodo-cytidine), 5- hydroxymethyl-cytidine (hm 5 C), 1-methyl-pseudoisocytidine, pyrrolo-cytidine, pyrrolo- 266 11979815v1 P-627574-PC pseudoisocytidine, 2-thio-cytidine (s 2 C), 2-thio-5-methyl-cy
  • a modified nucleobase is a modified adenine.
  • exemplary nucleobases and nucleosides having a modified adenine include 2-amino-purine, 2,6- diaminopurine, 2-amino-6-halo-purine (e.g., 2-amino-6-chloro-purine), 6-halo-purine (e.g., 6- chloro-purine), 2-amino-6-methyl-purine, 8-azido-adenosine, 7-deaza-adenine, 7-deaza-8-aza- adenine, 7-deaza-2-amino-purine, 7-deaza-8-aza-2-amino-purine, 7-deaza-2,6-diaminopurine, 7- deaza-8-aza-2,6-diaminopurine, 1-methyl-adenosine (m 1 A), 2-methyl-adenine (m 2 A), N6-methyl- adenine, 2-amin
  • a modified nucleobase is a modified guanine.
  • exemplary nucleobases and nucleosides having a modified guanine include inosine (I), 1-methyl-inosine (m 1 I), wyosine (imG), methylwyosine (mimG), 4-demethyl-wyosine (imG-14), isowyosine (imG2), wybutosine (yW), peroxywybutosine (o 2 yW), hydroxywybutosine (OHyW), undermodified hydroxywybutosine (OHyW*), 7-deaza-guanosine, queuosine (Q), 267 11979815v1 P-627574-PC epoxyqueuosine (oQ), galactosyl-queuosine (galQ), mannosyl-queuosine (manQ), 7-cyano-7- deaza-guanosine (pre
  • the nucleobase of the nucleotide can be independently selected from a purine, a pyrimidine, a purine or pyrimidine analog.
  • the nucleobase can each be independently selected from adenine, cytosine, guanine, uracil, or hypoxanthine.
  • the nucleobase can also include, for example, naturally-occurring and synthetic derivatives of a base, including pyrazolo[3,4-d]pyrimidines, 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo (e.g., 8-bromo), 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and
  • each letter refers to the representative base and/or derivatives thereof, e.g., A includes adenine or adenine analogs, e.g., 7-deaza adenine).
  • Modifications on the Internucleoside Linkage [0337]
  • the modified nucleotides which may be incorporated into a polynucleotide, primary construct, or RNA molecule, can be modified on the internucleoside linkage (e.g., phosphate backbone).
  • Backbone phosphate groups can be modified by replacing one or more of the oxygen atoms with a different substituent. Further, the modified nucleosides and nucleotides can include the wholesale replacement of an unmodified phosphate moiety with another internucleoside linkage as described herein.
  • modified phosphate groups include, but are not limited to, phosphorothioate, phosphoroselenates, boranophosphates, boranophosphate esters, hydrogen phosphonates, phosphoramidates, phosphorodiamidates, alkyl or aryl phosphonates, and phosphotriesters.
  • Phosphorodithioates have both non-linking oxygens replaced by sulfur.
  • the phosphate linker can also be modified by the replacement of a linking oxygen with nitrogen (bridged phosphoramidates), sulfur (bridged phosphorothioates), and carbon (bridged methylene-phosphonates).
  • the ⁇ -thio substituted phosphate moiety is provided to confer stability to RNA and DNA polymers through the unnatural phosphorothioate backbone linkages.
  • Phosphorothioate DNA and RNA have increased nuclease resistance and subsequently a longer half-life in a cellular environment.
  • Phosphorothioate linked polynucleotides, primary constructs, or modified RNA molecules are expected to also reduce the innate immune response through weaker binding/activation of cellular innate immune molecules.
  • a modified nucleoside includes an alpha-thio-nucleoside (e.g., 5'- O-(1-thiophosphate)-adenosine, 5'-O-(1-thiophosphate)-cytidine ( ⁇ -thio-cytidine), 5'-O-(1- thiophosphate)-guanosine, 5'-O-(1-thiophosphate)-uridine, or 5'-O-(1-thiophosphate)- pseudouridine).
  • alpha-thio-nucleoside e.g., 5'- O-(1-thiophosphate)-adenosine, 5'-O-(1-thiophosphate)-cytidine ( ⁇ -thio-cytidine), 5'-O-(1- thiophosphate)-guanosine, 5'-O-(1-thiophosphate)-uridine, or 5'-O-(1-thiophosphate)- pseudouridine
  • the polynucleotides, primary constructs, and modified RNA of the disclosure can include a combination of modifications to the sugar, the nucleobase, and/or the internucleoside linkage.
  • the purified preparation of RNA, oligoribonucleotide, or polyribonucleotide of the methods and compositions of the present disclosure comprises a combination of two or more of the above-described modifications.
  • the purified preparation of the RNA or oligoribonucleotide comprises a combination of three or more of the above-described modifications.
  • the purified preparation of the RNA 269 11979815v1 P-627574-PC or oligoribonucleotide comprises a combination of more than three of the above-described modifications.
  • the modified RNAs comprise in vitro-synthesized modified RNAs.
  • the present disclosure comprises one or more modified RNAs encoding an HSV glycoprotein.
  • the modified RNA comprises pseudouridine or pseudouridine family residues.
  • the modified RNAs of the present disclosure are capable of directing protein expression of HSV glycoproteins encoded thereon.
  • the present disclosure provides an in vitro-transcribed mRNA molecule encoding an HSV glycoprotein, comprising a pseudouridine.
  • the present disclosure provides a synthetic RNA molecule encoding an HSV glycoprotein, comprising a pseudouridine.
  • an in vitro-transcribed RNA molecule of the methods and compositions of the present disclosure is synthesized by T7 phage RNA polymerase.
  • the molecule is synthesized by SP6 phage RNA polymerase.
  • the molecule is synthesized by T3 phage RNA polymerase.
  • the molecule is synthesized by a polymerase selected from the above polymerases.
  • the RNA is synthesized chemically on a column similar to DNA.
  • the nucleoside that is modified in an RNA, oligoribonucleotide, or polyribonucleotide of the methods and compositions of the present disclosure is uridine (U).
  • the modified nucleoside is cytidine (C).
  • the modified nucleoside is adenine (A).
  • the modified nucleoside is guanine (G).
  • the RNA of the methods and compositions of the present disclosure further comprises a poly-A tail.
  • the RNA of the methods and compositions of the present disclosure does not comprise a poly-A tail.
  • the RNA of the methods and compositions of the present disclosure comprises an m7GpppG cap. In another embodiment, the RNA of the methods and compositions of the present disclosure does not comprise an m7GpppG cap. In another embodiment, the RNA of the methods and compositions of the present disclosure comprises a 3′- O-methyl-m7GpppG. In another embodiment, the RNA of methods and composition of the present 270 11979815v1 P-627574-PC disclosure comprise a non-reversible cap analog, which, in some embodiments, is added during transcription of the RNA. In another embodiment, the RNA of methods and composition of the present disclosure comprise an anti-reverse cap analog.
  • the RNA of the methods and compositions of the present disclosure further comprises a cap-independent translational enhancer.
  • the RNA of the methods and compositions of the present disclosure does not comprise a cap- independent translational enhancer.
  • the cap-independent translational enhancer is a tobacco etch virus (TEV) cap-independent translational enhancer.
  • the cap-independent translational enhancer is any other cap-independent translational enhancer known in the art.
  • “pseudouridine” refers to m 1 acp 3 ⁇ (1-methyl-3-(3-amino-5- carboxypropyl)pseudouridine. In another embodiment, the term refers to m 1 ⁇ (1- methylpseudouridine). In another embodiment, the term refers to ⁇ m (2′-O-methylpseudouridine. In another embodiment, the term refers to m 5 D (5-methyldihydrouridine). In another embodiment, the term refers to m 3 ⁇ (3-methylpseudouridine). In another embodiment, the modified nucleoside is 4' (pseudouridine).
  • a modified RNA comprises a modified nucleoside, which in some embodiments, comprises m 5 C, m5U, m 6 A, s 2 U, ⁇ ⁇ , 2'-O-methyl-U, 2’-O-methylpseudouridine, or a combination thereof.
  • the present disclosure provides a method for delivering a recombinant protein to a subject, the method comprising the step of contacting the subject with an RNA of the methods and compositions of the present disclosure, thereby delivering a recombinant protein to a subject.
  • a method of the present disclosure comprises increasing the number, percentage, or frequency of modified uridine nucleosides in the RNA molecule to decrease immunogenicity or increase efficiency of translation.
  • the number 271 11979815v1 P-627574-PC of modified uridine residues in an RNA, oligoribonucleotide, or polyribonucleotide molecule determines the magnitude of the effects observed in the present disclosure.
  • the fraction is 0.1%. In another embodiment, the fraction is 0.4%. In another embodiment, the fraction is 0.5%. In another embodiment, the fraction is 0.6%. In another embodiment, the fraction is 0.8%. In another embodiment, the fraction is 1%. In another embodiment, the fraction is 1.5%. In another embodiment, the fraction is 2%. In another embodiment, the fraction is 2.5%. In another embodiment, the fraction is 3%. In another embodiment, the fraction is 4%. In another embodiment, the fraction is 5%.
  • the fraction is 6%. In another embodiment, the fraction is 8%. In another embodiment, the fraction is 10%. In another embodiment, the fraction is 12%. In another embodiment, the fraction is 14%. In another embodiment, the fraction is 16%. In another embodiment, the fraction is 18%. In another embodiment, the fraction is 20%. In another embodiment, the fraction is 25%. In another embodiment, the fraction is 30%. In another embodiment, the fraction is 35%. In another embodiment, the fraction is 40%. In another embodiment, the fraction is 45%. In another embodiment, the fraction is 50%. In another embodiment, the fraction is 60%. In another embodiment, the fraction is 70%. In another embodiment, the fraction is 80%. In another embodiment, the fraction is 90%. In another embodiment, the fraction is 100%.
  • the fraction is less than 5%. In another embodiment, the fraction is less than 3%. In another embodiment, the fraction is less than 1%. In another embodiment, the fraction is less than 2%. In another embodiment, the fraction is less than 4%. In another embodiment, the fraction is less than 6%. In another embodiment, the fraction is less than 8%. In another embodiment, the fraction is less than 10%. In another embodiment, the fraction is less than 12%. In another embodiment, the fraction is less than 15%. In another embodiment, the fraction is less than 20%. In another embodiment, the fraction is less than 30%. In another embodiment, the fraction is less than 40%. In another embodiment, the fraction is less than 50%. In another embodiment, the fraction is less than 60%.
  • the fraction is less than 70% [0357] In another embodiment, 0.1% of the residues of a given uridine nucleotide are modified. In another embodiment, the fraction of the nucleotide is 0.2%. In another embodiment, the fraction 272 11979815v1 P-627574-PC is 0.3%. In another embodiment, the fraction is 0.4%. In another embodiment, the fraction is 0.5%. In another embodiment, the fraction is 0.6%. In another embodiment, the fraction is 0.8%. In another embodiment, the fraction is 1%. In another embodiment, the fraction is 1.5%. In another embodiment, the fraction is 2%. In another embodiment, the fraction is 2.5%. In another embodiment, the fraction is 3%. In another embodiment, the fraction is 4%. In another embodiment, the fraction is 5%.
  • the fraction is 6%. In another embodiment, the fraction is 8%. In another embodiment, the fraction is 10%. In another embodiment, the fraction is 12%. In another embodiment, the fraction is 14%. In another embodiment, the fraction is 16%. In another embodiment, the fraction is 18%. In another embodiment, the fraction is 20%. In another embodiment, the fraction is 25%. In another embodiment, the fraction is 30%. In another embodiment, the fraction is 35%. In another embodiment, the fraction is 40%. In another embodiment, the fraction is 45%. In another embodiment, the fraction is 50%. In another embodiment, the fraction is 60%. In another embodiment, the fraction is 70%. In another embodiment, the fraction is 80%. In another embodiment, the fraction is 90%. In another embodiment, the fraction is 100%.
  • the fraction of the given uridine nucleotide is less than 8%. In another embodiment, the fraction is less than 10%. In another embodiment, the fraction is less than 5%. In another embodiment, the fraction is less than 3%. In another embodiment, the fraction is less than 1%. In another embodiment, the fraction is less than 2%. In another embodiment, the fraction is less than 4%. In another embodiment, the fraction is less than 6%. In another embodiment, the fraction is less than 12%. In another embodiment, the fraction is less than 15%. In another embodiment, the fraction is less than 20%. In another embodiment, the fraction is less than 30%. In another embodiment, the fraction is less than 40%. In another embodiment, the fraction is less than 50%. In another embodiment, the fraction is less than 60%.
  • the fraction is less than 70%.
  • ribonucleotide refers to, in some embodiments, compounds comprising nucleotides in which the sugar moiety is ribose.
  • the term includes both RNA and RNA derivates in which the backbone is modified. Numerous RNA backbone modifications are known in the art and contemplated in the present disclosure.
  • modified RNA is a PNA (peptide nucleic acid). PNA contain peptide backbones and nucleotide bases and are able to bind, in another embodiment, to both DNA and RNA molecules.
  • nucleotide 273 11979815v1 P-627574-PC is modified by replacement of one or more phosphodiester bonds with a phosphorothioate bond.
  • the artificial nucleic acid contains any other variant of the phosphate backbone of native nucleic acids known in the art. Each nucleic acid derivative represents a separate embodiment of the present disclosure. [0360] Methods for production of nucleic acids having modified backbones are well known in the art, and are described, for example in U.S. Pat. Nos.5,723,335 and 5,663,153 issued to Hutcherson et al. and related PCT publication WO95/26204. Each method represents a separate embodiment of the present disclosure.
  • the nucleic acid of interest can be purified by any method known in the art, or any method to be developed, so long as the method of purification removes contaminants from the nucleic acid preparation and thereby substantially reduces the immunogenicity potential of the nucleic acid preparation.
  • the nucleic acid of interest is purified using high-performance liquid chromatography (HPLC).
  • HPLC high-performance liquid chromatography
  • the nucleic acid of interest is purified by contacting the nucleic acid of interest with the bacterial enzyme RNase III.
  • any method of nucleic acid purification that substantially reduces the immunogenicity of the nucleic acid preparation can be used.
  • Non-limiting examples of purification methods that can be used with the compositions and methods of the disclosure include liquid chromatography separation and enzyme digestion, each used alone or in any combination, simultaneously or in any order.
  • Non-limiting examples of liquid chromatography separation include HPLC and fast protein liquid chromatography (FPLC).
  • Materials useful in the HPLC and FPLC methods of the disclosure include, but are not limited to, cross-linked polystyrene/divinylbenzene (PS/DVB), PS/DVB-C18, PS/DVB-alkylated, Helix DNA columns (Varian), Eclipse dsDNA Analysis Columns (Agilent Technologies), Reverse-phase 5 (RPC-5) exchange material, DNAPac, ProSwift, and bio-inert UltiMate.RTM. 3000 Titanium columns (Dionex).
  • Enzymes useful in the enzyme digestion methods of the disclosure include any enzyme able to digest any contaminant in a nucleic acid preparation of the disclosure, such as, for example a dsRNA contaminant, and include but are not limited to, RNase III, RNase V1, Dicer, and Chipper (see Fruscoloni et al., 2002, PNAS 100:1639)
  • Non-limiting examples of assays for assessing the purity of the nucleic acid of interest include a dot-blot assay, a Northern blot assay, and a dendritic cell activation assay, as described elsewhere herein.
  • the modified RNA of the methods and compositions of the present disclosure is significantly less immunogenic than an unmodified in vitro-synthesized RNA 274 11979815v1 P-627574-PC molecule with the same sequence.
  • the modified RNA molecule is 2-fold less immunogenic than its unmodified counterpart.
  • immunogenicity is reduced by a 3-fold factor.
  • immunogenicity is reduced by a 5-fold factor.
  • immunogenicity is reduced by a 7-fold factor.
  • immunogenicity is reduced by a 10-fold factor.
  • immunogenicity is reduced by a 15-fold factor.
  • immunogenicity is reduced by a fold factor.
  • immunogenicity is reduced by a 50-fold factor. In another embodiment, immunogenicity is reduced by a 100-fold factor. In another embodiment, immunogenicity is reduced by a 200-fold factor. In another embodiment, immunogenicity is reduced by a 500-fold factor. In another embodiment, immunogenicity is reduced by a 1000-fold factor. In another embodiment, immunogenicity is reduced by a 2000-fold factor. In another embodiment, immunogenicity is reduced by another fold difference. [0363] In another embodiment, “significantly less immunogenic” refers to a detectable decrease in immunogenicity. In another embodiment, the term refers to a fold decrease in immunogenicity (e.g. 1 of the fold decreases enumerated above).
  • the term refers to a decrease such that an effective amount of the modified RNA can be administered without triggering a detectable immune response.
  • the term refers to a decrease such that the modified RNA can be repeatedly administered without eliciting an immune response sufficient to detectably reduce expression of the recombinant protein.
  • the decrease is such that the modified RNA can be repeatedly administered without eliciting an immune response sufficient to eliminate detectable expression of the recombinant protein.
  • the modified RNA of the methods and compositions of the present disclosure is translated in the cell more efficiently than an unmodified RNA molecule with the same sequence.
  • the modified RNA exhibits enhanced ability to be translated by a target cell.
  • translation is enhanced by a factor of 2-fold relative to its unmodified counterpart.
  • translation is enhanced by a 3-fold factor.
  • translation is enhanced by a 5-fold factor.
  • translation is enhanced by a 7-fold factor.
  • translation is enhanced by a 10- fold factor.
  • translation is enhanced by a 15-fold factor.
  • translation is enhanced by a 20-fold factor.
  • translation is 275 11979815v1 P-627574-PC enhanced by a 50-fold factor. In another embodiment, translation is enhanced by a 100-fold factor. In another embodiment, translation is enhanced by a 200-fold factor. In another embodiment, translation is enhanced by a 500-fold factor. In another embodiment, translation is enhanced by a 1000-fold factor. In another embodiment, translation is enhanced by a 2000-fold factor. In another embodiment, the factor is 10-1000-fold. In another embodiment, the factor is 10-100-fold. In another embodiment, the factor is 10-200-fold. In another embodiment, the factor is 10-300-fold. In another embodiment, the factor is 10-500-fold. In another embodiment, the factor is 20-1000- fold. In another embodiment, the factor is 30-1000-fold.
  • the factor is 50- 1000-fold. In another embodiment, the factor is 100-1000-fold. In another embodiment, the factor is 200-1000-fold. In another embodiment, translation is enhanced by any other significant amount or range of amounts. Each possibility represents a separate embodiment of the present disclosure.
  • Methods of determining translation efficiency are well known in the art, and include, e.g. measuring the activity of an encoded reporter protein (e.g., luciferase or renilla or green fluorescent protein [Wall A A, Phillips A M et al, Effective translation of the second cistron in two Drosophila dicistronic transcripts is determined by the absence of in-frame AUG codons in the first cistron.
  • an encoded reporter protein e.g., luciferase or renilla or green fluorescent protein
  • the target cell of the method of the present disclosure is a dendritic cell.
  • the target cell of the method of the present disclosure is a macrophage.
  • the target cell of the method of the present disclosure is a B cell.
  • the target cell of the method of the present disclosure is another antigen presenting cell.
  • the target cell of methods of the present disclosure is a mucosal cell.
  • the target cell of methods of the present disclosure is an epithelial cell.
  • the cell is a skin cell.
  • the cell is an epidermal cell.
  • the cell is a keratinocyte.
  • the cell is a Merkel cell, melanocyte or Langerhans cell. Each possibility represents a separate embodiment of the present disclosure.
  • C. Codon Optimization and GC Enrichment [0368] The present disclosure also provides codon optimized nucleotide sequences.
  • coding regions are codon-optimized for optimal expression in a subject to be treated using the RNA molecules described herein.
  • codon-optimization may be performed such that codons for which frequently occurring tRNAs are available are inserted in place of “rare codons.”
  • codon-optimization may include increasing guanosine/cytosine (G/C) content of a coding region of RNA described herein as compared to the G/C content of the corresponding coding sequence of a wild type RNA, wherein the amino acid sequence encoded by the RNA is preferably not modified compared to the amino acid sequence.
  • G/C guanosine/cytosine
  • a coding sequence (also referred to as a “coding region”) is codon optimized for expression in the subject to whom a composition (e.g., a pharmaceutical composition) is to be administered (e.g., a human).
  • sequences in such a polynucleotide e.g., a nucleotide sequence
  • sequences in such a polynucleotide may differ from wild type sequences encoding the relevant antigen, fragment or epitope thereof, even when the amino acid sequence of the antigen, fragment or epitope thereof is wild type.
  • strategies for codon optimization for expression in a relevant subject e.g., a human
  • a relevant subject e.g., a human
  • codon bias differs in codon usage between organisms
  • mRNA messenger RNA
  • tRNA transfer RNA
  • the predominance of selected tRNAs in a cell may generally be a reflection of the codons used most frequently in peptide synthesis. Accordingly, genes may be tailored for optimal gene expression in a given organism based on codon optimization.
  • Codon usage tables are available, for example, at the "Codon Usage Database” available at www.kazusa.orjp/codon/ and these tables may be adapted in a number of ways.
  • Computer algorithms for codon optimizing a particular sequence for expression in a particular subject or its cells are also available, such as Gene Forge (Aptagen; Jacobus, PA), are also available.
  • a polynucleotide e.g., a polyribonucleotide or a nucleotide sequence
  • a polynucleotide sequence of the present disclosure is codon optimized, wherein the codons in the polynucleotide (e.g., the polyribonucleotide) are adapted to human codon usage (herein referred to as “human codon optimized polynucleotide”).
  • a portion of a nucleotide sequence is codon optimized (e.g., a portion of or the portion encoding a glycoprotein or a portion of or the portion encoding a signal sequence).
  • the entire nucleotide sequence is codon optimized. Codons encoding the same amino acid occur at different frequencies in a subject, e.g., a human. Accordingly, in some embodiments, the coding sequence of a polynucleotide of the present disclosure is modified such that the frequency of the codons encoding the same amino acid corresponds to the naturally occurring frequency of that codon according to the human codon usage, e.g., as shown in Table 7.
  • the wild type coding sequence is preferably adapted in a way that the codon “GCC” is used with a frequency of 0.40, the codon “GCT” is used with a frequency of 0.28, the codon “GCA” is used with a frequency of 0.22 and the codon “GCG” is used with 30 a frequency of 0.10 etc. (see Table 7). Accordingly, in some embodiments, such a procedure (as exemplified for Ala) is applied for each amino acid encoded by the coding sequence of a polynucleotide to obtain sequences adapted to human codon usage. Table 7: Human codon usage with frequencies indicated for each amino acid.
  • a coding sequence may be optimized using a multiparametric optimization strategy.
  • optimization parameters may include parameters that influence protein expression, which can be, for example, impacted on a transcription level, an RNA level, and/or a translational level.
  • exemplary optimization parameters include, but are not limited to transcription-level parameters (including, e.g., GC content, consensus splice sites, cryptic splice sites, SD sequences, TATA boxes, termination signals, artificial recombination sites, and combinations thereof); RNA-level parameters (including, e.g., RNA instability motifs, 279 11979815v1 P-627574-PC ribosomal entry sites, repetitive sequences, and combinations thereof); translation-level parameters (including, e.g., codon usage, premature poly(A) sites, ribosomal entry sites, secondary structures, and combinations thereof); or combinations thereof.
  • transcription-level parameters including, e.g., GC content, consensus splice sites, cryptic splice sites, SD sequences, TATA boxes, termination signals, artificial recombination sites, and combinations thereof
  • RNA-level parameters including, e.g., RNA instability motifs, 279 11979815v1 P-627574-PC ribo
  • a coding sequence may be optimized by a GeneOptimizer algorithm as described in Fath et al. “Multiparameter RNA and Codon Optimization: A Standardized Tool to Assess and Enhance Autologous Mammalian Gene Expression” PLoS ONE 6(3): e17596; Rabb et al., which is incorporated herein by reference in its entirety, “The GeneOptimizer Algorithm: using a sliding window approach to cope with the vast sequence space in multiparameter DNA sequence optimization” Systems and Synthetic Biology (2010) 4:215-225; and Graft et al.
  • a coding sequence may be optimized by Eurofins’ adaption and optimization algorithm “GENEius” as described in Eurofins’ Application Notes: Eurofins’ adaption and optimization software “GENEius” in comparison to other optimization algorithms, the entire content of which is incorporated by reference for the purposes described herein.
  • a coding sequence utilized in accordance with the present disclosure has G/C content that is increased compared to a coding sequence for an HSV gC, gD, and/or gE (or immunogenic fragment thereof) construct described herein.
  • guanosine/cytidine (G/C) content of a coding region is modified relative to a comparable coding sequence for an HSV gC, gD, and/or gE (or immunogenic fragment thereof) construct described herein, but the amino acid sequence encoded by the nucleotide sequence is not modified.
  • sequences having an increased G (guanosine)/C (cytidine) content are more stable than sequences having an increased A (adenosine)/U (uridine) content.
  • G guanosine
  • C cytidine
  • A adenosine
  • U uridine
  • codons which contain A and/or U nucleosides can be modified by substituting these codons by other codons, which code 280 11979815v1 P-627574-PC for the same amino acids but contain no A and/or U or contain a lower content of A and/or U nucleosides.
  • G/C content of a coding region of a nucleotide sequence described herein is increased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G/C content of the coding region prior to codon optimization, e.g., of the wild type RNA.
  • G/C content of a coding region of a nucleotide sequence described herein is decreased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G/C content of the coding region prior to codon optimization, e.g., of the wild type RNA.
  • stability and translation efficiency of a nucleotide sequence may incorporate one or more elements established to contribute to stability and/or translation efficiency of the nucleotide sequence; exemplary such elements are described, for example, in PCT/EP2006/009448 incorporated herein by reference.
  • a nucleotide sequence may be modified within the coding region, i.e., the sequence encoding the expressed peptide or protein, without altering the sequence of the expressed peptide or protein, for example so as to increase the GC-content to increase RNA stability and/or to perform a codon optimization and, thus, enhance translation in cells.
  • Methods of Treatment and Uses of the Compositions [0379] In some embodiments, the present disclosure provides methods of inhibiting an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure.
  • the present disclosure provides methods of suppressing an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of preventing an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure. [0380] In some embodiments, an HSV oral mucosal infection is an HSV-1 infection.
  • an oral mucosal infection is an HSV-2 infection.
  • the present 281 11979815v1 P-627574-PC disclosure provides methods of treating an oral mucosal infection in a subject comprising administering a composition of the present disclosure.
  • the present disclosure provides methods of inhibiting an HSV-1 oral mucosal infection in a subject comprising administering a composition of the present disclosure.
  • the present disclosure provides methods of suppressing an HSV- 1 oral mucosal infection in a subject comprising administering a composition of the present disclosure.
  • the present disclosure provides methods of reducing the incidence of an HSV-1 oral mucosal infection in a subject comprising administering a composition of the present disclosure.
  • the present disclosure provides methods of preventing an HSV-1 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating an HSV-1 oral mucosal infection in a subject comprising administering a composition of the present disclosure. [0382] In some embodiments, the present disclosure provides methods of inhibiting an HSV-2 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of suppressing an HSV- 2 oral mucosal infection in a subject comprising administering a composition of the present disclosure.
  • the present disclosure provides methods of reducing the incidence of an HSV-2 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of preventing an HSV-2 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating an HSV-2 oral mucosal infection in a subject comprising administering a composition of the present disclosure. [0383] In some embodiments, the present disclosure provides methods of inhibiting HSV encephalitis in a subject comprising administering a composition of the present disclosure.
  • the present disclosure provides methods of suppressing HSV encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of HSV encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of preventing HSV encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present 282 11979815v1 P-627574-PC disclosure provides methods of treating HSV encephalitis in a subject comprising administering a composition of the present disclosure. [0384] In some embodiments, HSV encephalitis is an HSV-1 infection.
  • an oral mucosal infection is an HSV-2 infection.
  • the present disclosure provides methods of treating an oral mucosal infection in a subject comprising administering a composition of the present disclosure. [0385] In some embodiments, the present disclosure provides methods of inhibiting HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of suppressing HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure.
  • the present disclosure provides methods of preventing HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure. [0386] In some embodiments, the present disclosure provides methods of inhibiting HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of suppressing HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure.
  • the present disclosure provides methods of preventing HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure. [0387] In other embodiments, the present disclosure provides methods of preventing establishment of a latent HSV infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating a latent HSV infection in a subject comprising administering a composition of the present disclosure. In some embodiments, the latent HSV infection is an HSV-1 infection. In other embodiments, the latent HSV infection is an HSV-2 infection.
  • the latent HSV infection is in the dorsal root ganglia. In other embodiments, the latent HSV infection is in the trigeminal ganglia. In other embodiments, the latent HSV infection is in the brain. In other embodiments, the latent HSV infection is in the olfactory bulb. In other embodiments, the latent HSV infection is in any combination of dorsal root ganglia, trigeminal ganglia, brain, or olfactory bulb. [0389] In some embodiments, the latent HSV infection is detected in the dorsal root ganglia.
  • the latent HSV infection is detected in the trigeminal ganglia. In other embodiments, the latent HSV infection is detected in the brain. In other embodiments, the latent HSV infection is detected in the olfactory bulb. In other embodiments, the latent HSV infection is detected in any combination of dorsal root ganglia, trigeminal ganglia, brain, or olfactory bulb. In some embodiments, the latent HSV infection is an HSV-1 infection. In other embodiments, the latent HSV infection is an HSV-2 infection.
  • the present disclosure provides methods of suppressing or inhibiting an HSV infection of the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or a combination thereof in a subject comprising administering to said subject a composition of the present disclosure.
  • the present disclosure provides methods of treating or inhibiting an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof.
  • the HSV-2 protein comprises an HSV-2 glycoprotein, or an immunogenic fragment thereof.
  • the HSV-2 glycoprotein comprises an HSV-2 glycoprotein D (gD) or an immunogenic fragment thereof, an HSV glycoprotein C (gC) or an immunogenic fragment thereof, and an HSV glycoprotein E (gE) or an immunogenic fragment thereof.
  • said RNAs can be one or more RNAs comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6.
  • the present disclosure provides methods of inhibiting an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof.
  • the present disclosure provides methods of treating an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding 284 11979815v1 P-627574-PC Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof.
  • the present disclosure provides methods of suppressing an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof. In other embodiments, the present disclosure provides methods of reducing the incidence of an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof.
  • RNAs can be one or more RNAs comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6.
  • the present disclosure provides methods of vaccinating a subject against HSV and treating, impeding, inhibiting, reducing the incidence of, or suppressing an HSV infection or a symptom or manifestation thereof, comprising administration of a composition of the present disclosure.
  • the present disclosure provides a method for treating an HSV infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof.
  • the present disclosure provides a method for suppressing an HSV infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof.
  • the present disclosure provides a method for inhibiting an HSV infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof.
  • the present disclosure provides a method for reducing the incidence of HSV infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof. 285 11979815v1 P-627574-PC [0398]
  • the HSV infection is an HSV-1 infection.
  • the HSV infection is an HSV-2 infection.
  • the subject is administered HSV-1 glycoproteins, or immunogenic fragments thereof, for methods of treating, inhibiting, suppressing, etc. an HSV-1 infection.
  • the subject is administered HSV-2 glycoproteins, or immunogenic fragments thereof, for methods of treating, inhibiting, suppressing, etc. an HSV-2 infection.
  • the subject is administered HSV-1 glycoproteins, or immunogenic fragments thereof, for methods of treating, inhibiting, suppressing, etc. an HSV-1 infection, HSV-2 infection, or a combination thereof.
  • the subject is administered HSV-2 glycoproteins, or immunogenic fragments thereof, for methods of treating, inhibiting, suppressing, etc. an HSV-1 infection, HSV-2 infection, or a combination thereof.
  • HSV-1 glycoproteins e.g., gC1, gD1, gE1, immunogenic fragments thereof or a combination thereof
  • HSV-2 glycoproteins e.g., gC2, gD2, gE2, immunogenic fragments thereof, or a combination thereof
  • the present disclosure provides a method for treating, suppressing, inhibiting, or reducing the incidence of Herpes Simplex Virus 1 (HSV-1) infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV-1 glycoprotein or immunogenic fragment thereof.
  • HSV-1 Herpes Simplex Virus 1
  • the present disclosure provides a method for treating, suppressing, inhibiting, or reducing the incidence of Herpes Simplex Virus 2 (HSV-2) infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV-2 glycoprotein or immunogenic fragment thereof.
  • said contacting is via administration to said subject.
  • the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV gD or immunogenic fragment thereof; (b) an HSV gC or immunogenic fragment thereof as described herein; (c) an HSV gE or immunogenic fragment thereof as described herein, or a combination thereof.
  • the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV-2 gD or immunogenic fragment thereof; (b) an HSV-2 gC or immunogenic fragment thereof as described herein; and (c) an HSV-2 gE or immunogenic fragment thereof as described herein, or a combination thereof.
  • the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV-1 gD or immunogenic fragment thereof; (b) an HSV-1 gC or immunogenic fragment thereof as described herein; and (c) an HSV-1 gE or immunogenic fragment thereof as described herein, or a combination thereof.
  • the present disclosure provides a method of inducing an anti-HSV immune response in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV gD or immunogenic fragment thereof; (b) an HSV gC or immunogenic fragment thereof as described herein; (c) an HSV gE or immunogenic fragment thereof as described herein, or a combination thereof.
  • the present disclosure provides a method of inducing an anti-HSV immune response in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV-2 gD or immunogenic fragment thereof; (b) an HSV-2 gC or immunogenic fragment thereof as described herein; and (c) an HSV-2 gE or immunogenic fragment thereof as described herein, or a combination thereof.
  • the present disclosure provides a method of inducing an anti-HSV immune response in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV-1 gD or immunogenic fragment thereof; (b) an HSV-1 gC or immunogenic fragment thereof as described herein; and (c) an HSV-1 gE or immunogenic fragment thereof as described herein, or a combination thereof.
  • the present disclosure provides a method of inhibiting a primary HSV infection in a subject, the method comprising the step of administering to the subject a composition of the present disclosure.
  • the present disclosure provides a method of treating an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present 287 11979815v1 P-627574-PC disclosure provides a method of reducing the incidence of an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of inhibiting a flare following a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides methods of treating and/or suppressing a primary HSV infection and/or a secondary HSV infection.
  • a “primary” infection refers to a first-time infection.
  • a “secondary” infection refers to a recurrence of an HSV infection.
  • a “flare” or “recurrence” refers to reinfection of skin tissue following latent neuronal HSV infection.
  • the terms refer to reactivation of HSV after a latency period.
  • the terms refer to symptomatic HSV lesions following a non-symptomatic latency period.
  • the present disclosure provides a method of inhibiting spread of HSV. In some embodiments, the spread from DRG to skin is inhibited.
  • cell-to-cell spread of HSV is inhibited.
  • anterograde spread is inhibited.
  • retrograde spread is inhibited.
  • DDG refers, in some embodiments, to a neuronal cell body and in another embodiment, contain the neuron cell bodies of nerve fibers. In another embodiment, the term refers to any other definition of “DRG” used in the art.
  • spread of HSV to neural tissue is inhibited. [0413]
  • the present disclosure provides a method of inhibiting a recurrence following a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of preventing a recurrence following a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of inhibiting an HSV labialis following a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of preventing a recurrence of an HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a 288 11979815v1 P-627574-PC method of diminishing the severity of a recurrence of an HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of reducing the frequency of a recurrence of an HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides any of the described methods in an HIV-infected subject.
  • the present disclosure provides a method of treating HSV encephalitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of reducing the incidence of HSV encephalitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • HSV encephalitis refers, In some embodiments, to encephalitis caused by a Herpes Simplex Virus-1 (HSV).
  • HSV Herpes Simplex Virus-1
  • the term refers to encephalitis associated with HSV.
  • the term refers to any other type of HSV-mediated encephalitis known in the art.
  • the present disclosure provides a method of treating or reducing an HSV neonatal infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method for introducing an HSV glycoprotein to a cell of a subject, comprising contacting said cell with an in vitro-transcribed RNA molecule encoding the recombinant protein, wherein said in vitro-transcribed RNA molecule further comprises a modified nucleoside, thereby introducing said HSV glycoprotein, or immunogenic fragment thereof, into said cell of said subject.
  • the present disclosure provides a method for inducing a mammalian cell to produce an HSV glycoprotein, or immunogenic fragment thereof, comprising contacting said mammalian cell with an in vitro-synthesized RNA molecule encoding the HSV glycoprotein, or immunogenic fragment thereof, the in vitro-synthesized RNA molecule comprising a pseudouridine, thereby inducing said mammalian cell to produce said HSV glycoprotein, or immunogenic fragment thereof.
  • reference to HSV herein refers in some embodiments, to HSV- 1, while in another embodiment, to HSV-2, while in another embodiment, to HSV-1 and HSV-2.
  • HSV-1 refers to a Herpes Simplex Virus-1. In some embodiments, “HSV-1” refers to a HSV-1 strain. In another embodiment, the term refers to a KOS 289 11979815v1 P-627574-PC strain. In another embodiment, the term refers to an F strain. In another embodiment, the term refers to an NS strain. In another embodiment, the term refers to a CL101 strain. In another embodiment, the term refers to a “17” strain. In another embodiment, the term refers to a “17+syn” strain. In another embodiment, the term refers to a MacIntyre strain. In another embodiment, the term refers to an MP strain.
  • the term refers to an HF strain. In another embodiment, the term refers to any other HSV-1 strain known in the art.
  • “HSV-2” refers to a Herpes Simplex Virus-2. In some embodiments,“HSV-2” refers to a HSV-2 strain. In another embodiment, the term refers to an HSV-2333 strain. In another embodiment, the term refers to a 2.12 strain. In another embodiment, the term refers to an HG52 strain. In another embodiment, the term refers to an MS strain. In another embodiment, the term refers to a G strain. In another embodiment, the term refers to a 186 strain. In another embodiment, the term refers to any other HSV-2 strain known in the art.
  • the present disclosure provides a method of vaccinating a subject against an HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of suppressing an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of impeding an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of impeding a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of impeding neuronal HSV spread in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the terms “impeding an HSV infection” and “impeding a primary HSV infection” refer, in another embodiment, to decreasing the titer of infectious virus. In another embodiment, the terms refer to decreasing the extent of viral replication.
  • the present disclosure provides a method of treating an HSV- mediated herpetic ocular disease in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of reducing the incidence of an HSV-mediated herpetic ocular disease in a subject, the method comprising the step of administering to said subject a composition of the 290 11979815v1 P-627574-PC present disclosure.
  • the herpetic ocular disease comprises a corneal infection.
  • the present disclosure provides a method of treating an HSV-1 corneal infection or herpes keratitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of reducing the incidence of an HSV-1 corneal infection or herpes keratitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the HSV-1 corneal infection or herpes keratitis is an HSV-1 corneal infection or herpes keratitis.
  • the present disclosure provides a method of treating herpetic stomatitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of reducing the incidence of herpetic stomatitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the stomatitis is an HSV-1 stomatitis.
  • the present disclosure provides a method of treating, suppressing or inhibiting an HSV genital infection, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of treating, suppressing or inhibiting any manifestation of recurrent HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of reducing the incidence of an HSV-mediated genital ulcer disease in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of impeding establishment of a latent HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of preventing establishment of a latent HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of suppressing the establishment of a latent HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • establishment of a latent HSV infection in a subject is through HSV infection of the 291 11979815v1 P-627574-PC DRG.
  • establishment of a latent HSV infection in a subject is through HSV infection of the trigeminal nerve or trigeminal ganglia.
  • the present disclosure provides a method of treating, suppressing or inhibiting a genital herpes infection in a subject, comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of treating, suppressing or inhibiting an oral herpes infection in a subject, comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of treating or inhibiting an orolabial herpes infection in a subject, comprising the step of administering to said subject a composition of the present disclosure.
  • the HSV infection is on a lip of the subject.
  • the present disclosure provides a method of reducing the incidence of an HSV-mediated encephalitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the herpes-mediated encephalitis treated or prevented by a method of the present disclosure is a focal herpes encephalitis.
  • the herpes- mediated encephalitis is a neonatal herpes encephalitis.
  • the herpes- mediated encephalitis is any other type of herpes-mediated encephalitis known in the art.
  • the present disclosure provides a method of treating or reducing the incidence of a disease, disorder, or symptom associated with or secondary to an HSV-mediated encephalitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of treating, reducing the pathogenesis of, ameliorating the symptoms of, ameliorating the secondary symptoms of, reducing the incidence of, prolonging the latency to a relapse of an HSV infection in a subject, comprising the step of administering to the subject a composition of the present disclosure.
  • the present disclosure provides a method of protecting a subject against formation of a zosteriform lesion or an analogous outbreak in a human subject.
  • the present disclosure provides a method of inhibiting the formation of an HSV zosteriform lesion or an analogous outbreak in a human subject.
  • Zosteriform refers, in some embodiments, to skin lesions characteristic of an HSV infection, particularly during reactivation infection, which, in some embodiments, begin as a rash 292 11979815v1 P-627574-PC and follow a distribution near dermatomes, commonly occurring in a strip or belt-like pattern. In some embodiments, the rash evolves into vesicles or small blisters filled with serous fluid. In some embodiments, zosteriform lesions form in mice as a result of contact with HSV. In another embodiment, zosteriform lesions form in humans as a result of contact with HSV.
  • Zerosteriform spread refers, in some embodiments, to an HSV infection that spreads from the ganglia to secondary skin sites within the dermatome. In another embodiment, the term refers to spread within the same dermatome as the initial site of infection. In another embodiment, the term refers to any other definition of “zosteriform spread” known in the art. “Outbreak”, in another embodiment, refers to a sudden increase in symptoms of a disease or in the spread or prevalence of a disease, and in some embodiments, refers to a sudden increase in zosteriform lesions, while in another embodiment, “outbreak” refers to a sudden eruption of zosteriform lesions.
  • the present disclosure provides a method of impeding the formation of a dermatome lesion or an analogous condition in a subject.
  • dermatome lesions form as a result of contact with HSV.
  • dermatome lesions most often develop when the virus reactivates from latency in the ganglia and in some embodiments, spreads down nerves, in some embodiments, causing a recurrent infection.
  • the methods of the present disclosure may be used to treat, inhibit, suppress, etc. an HSV infection or primary or secondary symptoms related to such an infection following exposure of the subject to HSV.
  • the subject has been infected with HSV before vaccination.
  • the subject is at risk for HSV infection.
  • vaccination by a method of the present disclosure is efficacious in treating, inhibiting, suppressing, etc. an HSV infection or primary or secondary symptoms related to such an infection.
  • “treating” refers to either therapeutic treatment or prophylactic or preventative measures, wherein the object is to prevent or lessen the targeted pathologic condition or disorder as described hereinabove.
  • treating may include directly affecting or curing, suppressing, inhibiting, preventing, reducing the severity of, delaying the onset of, reducing symptoms associated with the disease, disorder or condition, or a combination thereof.
  • “treating” refers inter alia to delaying progression, expediting remission, inducing remission, augmenting remission, speeding recovery, increasing efficacy of or decreasing resistance to alternative therapeutics, or a combination thereof.
  • “preventing” refers, inter alia, to delaying the onset of symptoms, preventing 293 11979815v1 P-627574-PC relapse to a disease, decreasing the number or frequency of relapse episodes, increasing latency between symptomatic episodes, or a combination thereof.
  • “suppressing” or “inhibiting”, refers inter alia to reducing the severity of symptoms, reducing the severity of an acute episode, reducing the number of symptoms, reducing the incidence of disease-related symptoms, reducing the latency of symptoms, ameliorating symptoms, reducing secondary symptoms, reducing secondary infections, prolonging patient survival, or a combination thereof.
  • the compositions and methods of the present disclosure are effective in lowering HSV acquisition rates, duration of HSV infection, frequency of HSV reactivation, or a combination thereof.
  • the compositions and methods of the present disclosure are effective in treating or inhibiting genital ulcer disease, which in some embodiments, entails decreasing the severity or frequency of HSV genital ulcer disease.
  • compositions and methods of the present disclosure block immune evasion from complement.
  • vaccination with RNA-encoded HSV subunits, or immunogenic fragments thereof may produce high titers of neutralizing antibodies or potent T-cell responses; however, upon subsequent infection, HSV immune evasion molecules may block the activities of antibodies or T cells, thereby reducing composition efficacy.
  • the compositions and methods of the present disclosure incorporate strategies to block virus mediated immune evasion by, in some embodiments, enhancing the effectiveness of e.g. a gD-1 subunit, or immunogenic fragment thereof, composition using gC-1, or immunogenic fragments thereof, to prevent immune evasion from complement.
  • compositions and methods of the present disclosure are useful for preventing or inhibiting recurrent HSV infections.
  • antibodies to e.g. gC-1 block domains involved in immune evasion, which enhances complement activity, improves neutralizing activity of anti- gD-1 IgG, increases antibody- and complement-dependent cellular cytotoxicity, and augments complement-mediated neutralization and lysis of infected cells.
  • symptoms are primary, while in another embodiment, symptoms are secondary.
  • “primary” refers to a symptom that is a direct result of the subject viral infection
  • “secondary” refers to a symptom that is derived from or consequent to a primary cause.
  • the compositions and strains 294 11979815v1 P-627574-PC for use in the present disclosure treat primary or secondary symptoms or secondary complications related to HSV infection.
  • symptoms may be any manifestation of an HSV infection, comprising blisters, ulcerations, or lesions on the urethra, cervix, upper thigh, and/or anus in women and on the penis, urethra, scrotum, upper thigh, and anus in men, inflammation, swelling, fever, flu-like symptoms, sore mouth, sore throat, pharyngitis, pain, blisters on tongue, mouth or lips, ulcers, cold sores, neck pain, enlarged lymph nodes, reddening, bleeding, itching, dysuria, headache, muscle pain, etc., or a combination thereof.
  • the disease, disorder, or symptom is fever.
  • the disease, disorder, or symptom is headache. In another embodiment, the disease, disorder, or symptom is stiff neck. In another embodiment, the disease, disorder, or symptom is seizures. In another embodiment, the disease, disorder, or symptom is partial paralysis. In another embodiment, the disease, disorder, or symptom is stupor. In another embodiment, the disease, disorder, or symptom is coma. In another embodiment, the disease, disorder, or symptom is any other disease, disorder, or symptom known in the art that is associated with or secondary to a herpes-mediated encephalitis. [0446] Methods of determining the presence and severity of herpes-mediated encephalitis are well known in the art, and are described, for example, in Bonkowsky JL et al.
  • the present disclosure provides a method of treating or reducing the incidence of a disease, disorder, or symptom associated with an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the disease, disorder, or symptom secondary to an HSV infection is oral lesions.
  • the disease, disorder, or symptom is genital lesions. In another embodiment, the disease, disorder, or symptom is oral ulcers. In another embodiment, the disease, disorder, or symptom is genital ulcers. In another embodiment, the disease, disorder, or symptom is fever. In another embodiment, the disease, disorder, or symptom is headache. In another embodiment, the disease, disorder, or symptom is muscle ache. In another embodiment, 295 11979815v1 P-627574-PC the disease, disorder, or symptom is swollen glands in the groin area. In another embodiment, the disease, disorder, or symptom is painful urination. In another embodiment, the disease, disorder, or symptom is vaginal discharge.
  • the disease, disorder, or symptom is blistering. In another embodiment, the disease, disorder, or symptom is flu-like malaise. In another embodiment, the disease, disorder, or symptom is keratitis. In another embodiment, the disease, disorder, or symptom is herpetic whitlow. In another embodiment, the disease, disorder, or symptom is Bell's palsy. In another embodiment, the disease, disorder, or symptom is herpetic erythema multiforme. In another embodiment, the disease, disorder, or symptom is a lower back symptom (e.g. numbness, tingling of the buttocks or the area around the anus, urinary retention, constipation, and impotence).
  • a lower back symptom e.g. numbness, tingling of the buttocks or the area around the anus, urinary retention, constipation, and impotence.
  • the disease, disorder, or symptom is a localized eczema herpeticum. In another embodiment, the disease, disorder, or symptom is a disseminated eczema herpeticum. In another embodiment, the disease, disorder, or symptom is a herpes gladiatorum. In another embodiment, the disease, disorder, or symptom is a herpetic sycosis. In another embodiment, the disease, disorder, or symptom is an esophageal symptom (e.g. difficulty swallowing or burning, squeezing throat pain while swallowing, weight loss, pain in or behind the upper chest while swallowing).
  • esophageal symptom e.g. difficulty swallowing or burning, squeezing throat pain while swallowing, weight loss, pain in or behind the upper chest while swallowing.
  • the disease, disorder, or symptom is any other disease, disorder, or symptom that is known in the art.
  • Each disease, disorder, and symptom represents a separate embodiment of the present disclosure.
  • the compositions and methods of the instant disclosure treat, suppress, inhibit, or reduce the incidence of the infection itself, while in another embodiment, the compositions and methods of the instant disclosure treat, suppress, inhibit, or reduce the incidence of primary symptoms of the infection, while in another embodiment, the compositions and methods of the instant disclosure treat, suppress, inhibit, or reduce the incidence of secondary symptoms of the infection. It is to be understood that the compositions and methods of the instant disclosure may affect any combination of the infection, the primary symptoms caused by the infection, and secondary symptoms related to the infection.
  • the HSV infection that is treated or ameliorated by methods and compositions of the present disclosure is, in another embodiment, a genital HSV infection.
  • the HSV infection is an oral HSV infection.
  • the HSV infection is an ocular HSV infection.
  • the HSV infection is a dermatologic HSV infection.
  • the HSV infection is a neurological HSV infection.
  • the HSV infection is a neuronal HSV infection. 296 11979815v1 P-627574-PC [0451]
  • the present disclosure provides a method of reducing the incidence of a disseminated HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of reducing the incidence of a neonatal HSV infection in an offspring of a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of reducing a transmission of an HSV infection from a subject to an offspring thereof, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the offspring is an infant.
  • the transmission that is reduced or inhibited is transmission during birth.
  • transmission during breastfeeding is reduced or inhibited.
  • the transmission that is reduced or inhibited is any other type of parent-to-offspring transmission known in the art.
  • the present disclosure provides a method of reducing a severity of a neonatal HSV infection in an offspring of a subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition comprising: (a) an RNA encoding HSV gC protein or fragment thereof; (b) an RNA encoding HSV gE protein or fragment thereof; and (c) an adjuvant.
  • the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition comprising: (a) an RNA encoding HSV gC protein or fragment thereof, wherein said fragment comprises either a C3b-binding domain thereof, a properdin interfering domain thereof, a C5 interfering domain thereof, or a fragment of said C3b-binding domain, properdin interfering domain, or C5-interfering domain; (b) an RNA encoding HSV gE protein or fragment thereof, wherein said fragment comprises AA 24-409, 24-405, or a fragment thereof; and (c) an adjuvant.
  • the present disclosure provides a method of treating an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of suppressing an HSV infection in a subject infected with HIV, the method 297 11979815v1 P-627574-PC comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of inhibiting an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of reducing the incidence of an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of preventing an HIV infection, the method comprising the step of administering to said subject an HSV composition of the present disclosure.
  • HSV infection increases the risk of HIV infection, and protection against HSV infection decreases the risk of HIV infection.
  • the present disclosure provides a method of decreasing the risk of an HIV infection, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the composition for use in the methods of the present disclosure elicits an immune response against HSV.
  • the composition for use in the methods of the present disclosure elicits an immune response against HSV-1.
  • the composition for use in the methods of the present disclosure elicits an immune response against HSV-2.
  • the composition comprises RNAs encoding gD and gC proteins, or immunogenic fragments thereof.
  • the composition comprises RNAs encoding gE and gD proteins, or immunogenic fragments thereof.
  • the composition comprises RNAs encoding gC and gE proteins, or immunogenic fragments thereof.
  • the composition comprises RNAs encoding gE, gD, and gC proteins, or immunogenic fragments thereof. In another embodiment, the composition comprises RNAs encoding gE, gD, or gC protein, or immunogenic fragments thereof. In another embodiment, the proteins encoded by the RNAs are HSV-1 proteins, or immunogenic fragments thereof. In another embodiment, the proteins encoded by the RNAs are HSV-2 proteins, or immunogenic fragments thereof. In another embodiment, the proteins encoded by the RNAs comprise both HSV-1 and HSV-2 proteins, or immunogenic fragments thereof.
  • a subject according to any of the embodiments described herein may be a subject infected with, or in another embodiment, susceptible to infection with HSV.
  • a subject may be infected with, or in another embodiment, susceptible to infection with at least one other pathogen.
  • a subject may be immunocompromised.
  • the subject is 298 11979815v1 P-627574-PC infected by HSV, while in another embodiment, the subject is at risk for infection by HSV, which in some embodiments, is a subject who is a neonate, in another embodiment, immunocompromised, in another embodiment, elderly, and in another embodiment, an immunocompromised neonate or an immunocompromised elderly subject.
  • the compositions of the present disclosure and their related uses may suppress, inhibit, prevent or treat an HIV infection in a subject.
  • the compositions of the present disclosure and their related uses may treat secondary complications of HIV infection, which in some embodiments, are opportunistic infections, neoplasms, neurologic abnormalities, or progressive immunologic deterioration.
  • the methods comprise treating acquired immunodeficiency syndrome (AIDS).
  • the methods comprise treating a decline in the number of CD4 + T lymphocytes.
  • the present disclosure provides a method of reducing HIV-1 transmission to an offspring, the method comprising the step of administering to a subject a composition of the present disclosure.
  • HSV-2 infection increases HIV-1 viral shedding in genital secretions (Nagot N et al., Reduction of HIV-1 RNA levels with therapy to suppress herpes simplex virus. N Engl J Med. 2007 Feb 22; 356(8):790-9).
  • methods of the present disclosure of inhibiting HSV-2 infection are also efficacious for reducing HIV-1 transmission to an offspring.
  • the mutant HSV strain is an HSV-1 strain.
  • the mutant HSV strain is an HSV-2 strain.
  • the present disclosure provides a method of reducing HIV-1 transmission to a sexual partner, the method comprising the step of administering to a subject a composition of the present disclosure.
  • the mutant HSV strain is an HSV-1 strain.
  • the mutant HSV strain is an HSV-2 strain.
  • the present disclosure provides a method of reducing susceptibility to HIV-1, the method comprising the step of administering to a subject a composition of the present disclosure.
  • HSV-2 infection increases HIV-1 replication (Ouedraogo A et al., Impact of suppressive herpes therapy on genital HIV-1 RNA among women taking antiretroviral therapy: a randomized controlled trial. AIDS. 2006 Nov 28;20(18):2305-13).
  • methods of the present disclosure of inhibiting HSV-2 infection are also efficacious for reducing 299 11979815v1 P-627574-PC susceptibility to HIV-1.
  • the mutant HSV strain is an HSV-1 strain.
  • the mutant HSV strain is an HSV-2 strain.
  • the present disclosure provides a method of inhibiting a primary HSV infection in an HIV-infected subject, comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of treating or reducing the incidence of an HSV infection in an HIV-infected subject, comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method of inhibiting a flare, recurrence, or HSV labialis following a primary HSV infection in an HIV-infected subject, the method comprising the step of administering to said subject a composition of the present disclosure.
  • the present disclosure provides a method for inducing an immune response in a subject, the method comprising the step of administering to said subject a nucleoside RNA composition of the present disclosure.
  • the immune response comprises a CD4 immune response.
  • the immune response comprises a CD8 immune response.
  • the immune response comprises a T follicular helper cell immune response.
  • the immune response comprises a germinal center B cell immune response.
  • the immune response comprises an IgG antibody response to gC2, gD2, gE2, immunogenic fragments thereof, or a combination thereof.
  • the present disclosure provides a method of treating a Herpes Simplex Virus (HSV) infection in a subject, the method comprising the step of intramuscularly administering to said subject a nucleoside RNA composition of the present disclosure.
  • the disclosure provides a method of suppressing, inhibiting, or reducing the incidence of a Herpes Simplex Virus (HSV) infection in a subject, the method comprising the step of intramuscularly administering to said subject a nucleoside RNA composition of the present disclosure.
  • the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence a Herpes Simplex Virus (HSV) infection in a subject, the method comprising the step of administering to said subject a nucleoside RNA composition of the present disclosure and further comprising administering to said subject one or more polypeptide compositions comprising the proteins or immunogenic fragments thereof that are encoded by the 300 11979815v1 P-627574-PC nucleoside RNA as described herein.
  • HSV Herpes Simplex Virus
  • the polypeptide composition comprises (a) a composition comprising HSV gD or an immunogenic fragment thereof, (b) a composition comprising HSV gC or an immunogenic fragment thereof, and (c) a composition comprising HSV gE or an immunogenic fragment thereof.
  • the disclosure provides a method of of a Herpes Simplex Virus (HSV) infection in a subject, the method comprising the step of intramuscularly administering to said subject a nucleoside RNA composition of the present disclosure.
  • HSV Herpes Simplex Virus
  • the present disclosure provides a method as described herein wherein: the HSV-2 gC or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 28, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 241, or SEQ ID NO: 242; the HSV-2 gD or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 110, SEQ ID NO: 234, or SEQ ID NO: 120; and/or the HSV-2 gE or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 17 or SEQ ID NO: 18.
  • the present disclosure provides a method as described herein wherein: the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 25, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239, or SEQ ID NO: 246; the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 23, SEQ ID NO: 135, SEQ ID NO: 23, S
  • the present disclosure provides a method as described herein wherein: the HSV-2 gC or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 224; the HSV-2 gD or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 5; and/or the HSV-2 gE or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 17.
  • the present disclosure provides a method as described herein wherein: the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 246; the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 247; and/or the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 145.
  • the present disclosure provides a method as described herein wherein: the HSV-1 gC or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 240; the HSV-1 gD or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 134; and/or the HSV-1 gE or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 248, or SEQ ID NO: 140.
  • the present disclosure provides a method as described herein wherein: the RNA encoding HSV-1 gC or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 24; the RNA encoding HSV-1 gD or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 22; and/or the RNA encoding HSV-1 gE or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 26. 302 11979815v1 P-627574-PC [0475]
  • the present disclosure provides a method as described herein wherein the signal sequence is any signal sequence listed in Table 3.
  • the signal sequence is a viral signal sequence.
  • the viral signal sequence is an HSV signal sequence.
  • the HSV signal sequence is an HSV gD signal sequence.
  • the HSV gD signal sequence is an HSV-1 gD signal sequence.
  • the HSV-1 gD signal sequence encodes a polypeptide comprising a sequence at least 85% identical to SEQ ID NO: 34 or SEQ ID NO: 35.
  • the HSV-1 gD signal sequence encodes a polypeptide comprising or consisting of a sequence at least 85% identical to SEQ ID NO: 35.
  • the HSV gD signal sequence is an HSV-2 gD signal sequence.
  • the HSV-2 gD signal sequence encodes a polypeptide comprising a sequence at least 85% identical to SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, or SEQ ID NO: 33.
  • the HSV-2 gD signal sequence encodes a polypeptide comprising or consisting of a sequence at least 85% identical to SEQ ID NO: 29 or SEQ ID NO: 31.
  • the signal sequence is a codon optimized signal sequence.
  • the method comprises administering to said subject: a composition comprising RNA at least 85% identical to SEQ ID NO: 246; a composition comprising RNA at least 85% identical to SEQ ID NO: 247; and a composition comprising RNA at least 85% identical to SEQ ID NO: 145.
  • compositions of the present disclosure can be, in another embodiment, administered to a subject by any method known to a person skilled in the art, such as parenterally, paracancerally, transmucosally, transdermally, intramuscularly, intravenously, intra-dermally, subcutaneously, intra-peritonealy, intra-ventricularly, intra-cranially, intra-vaginally, intra-nasally, intra-tumorally, or topically.
  • administering in another embodiment, refers to directly introducing into a subject by injection or other means a composition of the present disclosure.
  • “administering” refers to contacting a cell of the subject’s immune system with a composition or RNA encoding HSV protein or mixture thereof. 303 11979815v1 P-627574-PC [0489]
  • the compositions are administered orally, and are thus formulated in a form suitable for oral administration, i.e. as a solid or a liquid preparation.
  • Suitable solid oral formulations include tablets, capsules, pills, granules, pellets and the like.
  • Suitable liquid oral formulations include solutions, suspensions, dispersions, emulsions, oils and the like.
  • the active ingredient is formulated in a capsule.
  • the compositions of the present disclosure comprise, in addition to the active compound and the inert carrier or diluent, a hard gelating capsule.
  • the pharmaceutical compositions are administered by intravenous, intra-arterial, or intramuscular injection of a liquid preparation. Suitable liquid formulations include solutions, suspensions, dispersions, emulsions, oils and the like.
  • the pharmaceutical compositions are administered intravenously and are thus formulated in a form suitable for intravenous administration.
  • the pharmaceutical compositions are administered intra-arterially and are thus formulated in a form suitable for intra-arterial administration.
  • the pharmaceutical compositions are administered intramuscularly and are thus formulated in a form suitable for intramuscular administration.
  • the pharmaceutical compositions are administered topically to body surfaces and are thus formulated in a form suitable for topical administration. Suitable topical formulations include gels, ointments, creams, lotions, drops and the like.
  • the compositions or their physiologically tolerated derivatives are prepared and applied as solutions, suspensions, or emulsions in a physiologically acceptable diluent with or without a pharmaceutical carrier.
  • the composition is administered as a suppository, for example a rectal suppository or a urethral suppository.
  • the pharmaceutical composition is administered by subcutaneous implantation of a pellet.
  • the pellet provides for controlled release of agent over a period of time.
  • pharmaceutical compositions are administered intramuscularly, subcutaneously or intradermally.
  • Effective dosage refers, in another embodiment, to an amount sufficient to exert a therapeutic effect. In another embodiment, the term refers to an amount sufficient to elicit expression of a detectable amount of the encoded protein. Each possibility represents a separate embodiment of the present disclosure. 304 11979815v1 P-627574-PC [0495] Methods for measuring the dose of an RNA encoding an HSV glycoprotein, or an immunogenic fragment thereof (e.g.
  • any of the HSV compositions of and for use in the methods of this disclosure will comprise an RNA encoding HSV protein or an immunogenic fragment thereof or combination of RNAs encoding HSV proteins or immunogenic fragments thereof, of the present disclosure, in any form or embodiment as described herein.
  • any of the compositions of and for use in the methods will consist of an RNA encoding an HSV protein, or immunogenic fragment thereof, or combination of RNA encoding HSV proteins, or immunogenic fragments thereof, of the present disclosure, in any form or embodiment as described herein.
  • compositions of this disclosure will consist essentially of an RNA encoding an HSV protein, or immunogenic fragment thereof, or combination of RNAs encoding HSV proteins, or an immunogenic fragments thereof, of the present disclosure, in any form or embodiment as described herein.
  • the term “comprise” refers to the inclusion of RNA encoding other HSV proteins, or immunogenic fragments thereof, as well as inclusion of RNA encoding other proteins that may be known in the art.
  • the term “consisting essentially of” refers to a composition, which has the RNA encoding a specific HSV protein or fragment thereof.
  • the term “consisting” refers to a composition having an RNA encoding particular HSV protein or fragment or combination of RNAs encoding HSV proteins or fragments of the present disclosure, in any form or embodiment as described herein.
  • the present disclosure provides a composition for treating HSV-1 or a symptom or manifestation thereof, the composition comprising an RNA of the present disclosure.
  • the present disclosure provides a composition for treating HSV-2 or a symptom or manifestation thereof, the composition comprising an RNA of the present disclosure.
  • compositions, and methods of the present disclosure may be used in non-HSV herpesvirus as well, which in some embodiments, proteins gD, gE, or gC proteins that are, in some embodiments, 70% homologous, in another embodiment, 80% homologous, in 305 11979815v1 P-627574-PC another embodiment, 85% homologous, in another embodiment, 90% homologous, in another embodiment, 95% homologous, in another embodiment, 98% homologous, and in another embodiment, 100% homologous to the gD, gE, or gC proteins of HSV-1, or in another embodiment, of HSV-2.
  • proteins gD, gE, or gC proteins that are, in some embodiments, 70% homologous, in another embodiment, 80% homologous, in 305 11979815v1 P-627574-PC another embodiment, 85% homologous, in another embodiment, 90% homologous, in another embodiment, 95% homologous, in another embodiment, 98% homologous, and in another
  • non-HSV herpesvirus comprise Varicella Zoster Virus (VZV), Epstein-Barr virus (EBV), EBNA, cytomegalovirus (CMV), and human herpesvirus-6 (HHV-6).
  • VZV Varicella Zoster Virus
  • EBV Epstein-Barr virus
  • CMV cytomegalovirus
  • HHV-6 human herpesvirus-6
  • the dosage is a daily dose. In another embodiment, the dosage is a weekly dose. In another embodiment, the dosage is a monthly dose. In another embodiment, the dosage is an annual dose. In another embodiment, the dose is one is a series of a defined number of doses. In another embodiment, the dose is a one-time dose.
  • any of the booster doses described hereinabove is administered following a priming dose comprising one or modified more RNAs encoding HSV-1 proteins or immunogenic fragments thereof. In another embodiment, any of the booster doses described hereinabove is administered following a priming vaccination comprising one or more modified more RNAs encoding HSV-2 proteins or immunogenic fragments thereof.
  • a subject is immunized with a single administration of the composition. In another embodiment, a subject is immunized with a single dose. In another embodiment, a subject is immunized with two doses. In another embodiment, a subject is immunized with three doses. In another embodiment, a subject is immunized with four doses. In another embodiment, a subject is immunized with five doses. [0504] In some embodiments, all the components of the composition are provided in equal concentrations. According to this aspect and in some embodiments, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are provided in a ratio of 1:1:1.
  • RNAs encoding gC, gD, and gE, or immunogenic fragments thereof are provided in a ratio of 5:2:5. In another embodiment, RNAs encoding gC and gD, or immunogenic fragments thereof, are provided in a ratio of 1:1. In another embodiment, RNAs encoding gC and gE, or immunogenic 306 11979815v1 P-627574-PC fragments thereof, are provided in a ratio of 1:1. In another embodiment, RNAs encoding gD and gE, or immunogenic fragments thereof, are provided in a ratio of 1:1.
  • RNAs encoding gC, gD, gE, immunogenic fragments thereof, or a combination thereof, or combined with other HSV glycoproteins are administered in a single composition at the same site and by the same route, while in another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered in separate compositions at separate sites but by the same route of administration, or in another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered in separate compositions at separate sites and by different routes of administration, or in another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered in separate compositions at the same site and by different routes of administration (e.g.
  • the methods of the present disclosure include a one-time or single administration of compositions comprising one or more nucleoside RNAs of the present disclosure.
  • the methods of the present disclosure include administration of compositions comprising one or more nucleoside RNAs in a prime and boost approach.
  • the methods of the present disclosure further comprise the step of administering to said subject one or more additional administrations of said nucleoside RNA composition subsequent to the first administration.
  • the methods of the present disclosure comprise administering a composition comprising one or more nucleoside RNAs encoding one or more HSV glycoproteins, or immunogenic fragments thereof, as a first administration and a composition comprising one or more HSV glycoproteins, or immunogenic fragments thereof, as a second or subsequent administration.
  • the HSV glycoproteins, or immunogenic fragments thereof, encoded by an RNA in the first (or prime) administration are the same glycoproteins, or fragments thereof, in the second or subsequent (or boost) administration.
  • a composition comprising one or more HSV glycoproteins, or immunogenic fragments thereof is administered as a first administration, and a composition comprising one or more nucleoside RNAs encoding one or more HSV glycoproteins, or immunogenic fragments thereof, is administered as a second or subsequent administration.
  • RNAs encoding gC, gD, and gE, or immunogenic fragments thereof are administered simultaneously followed by a booster dose of RNA encoding gD without 307 11979815v1 P-627574-PC RNAs encoding gC or gE, or immunogenic fragments thereof.
  • RNAs encoding gC, gD, and gE, or immunogenic fragments thereof are administered simultaneously followed by a booster dose of RNA encoding gC without RNAs encoding gD or gE, or immunogenic fragments thereof,.
  • RNAs encoding gC, gD, and gE, or immunogenic fragments thereof are administered simultaneously followed by a booster dose of RNA encoding gE without RNAs encoding gD or gC, or immunogenic fragments thereof,.
  • RNAs encoding gC, gD, and gE, or immunogenic fragments thereof are administered simultaneously followed by a booster dose of RNAs encoding gC and gD, or immunogenic fragments thereof, without RNAs encoding gE, or immunogenic fragments thereof,.
  • RNAs encoding gC, gD, and gE, or immunogenic fragments thereof are administered simultaneously followed by a booster dose of RNAs encoding gC and gE, or immunogenic fragments thereof, without RNAs encoding gD, or immunogenic fragments thereof,.
  • RNAs encoding gC, gD, and gE, or immunogenic fragments thereof are administered simultaneously followed by a booster dose of RNAs encoding gD and gE, or immunogenic fragments thereof, without RNAs encoding gE, or immunogenic fragments thereof,.
  • the booster administration is performed at the same site and by the same mode of administration as the priming administration.
  • the booster administration is performed at a different site from the priming administration but by the same mode of administration as the priming administration.
  • the booster administration is performed at the same site but by different mode of administration than priming administration.
  • the booster administration is performed at a different site and by different mode of administration than priming administration.
  • the modified RNA induces a detectably lower innate immune response than the same quantity of unmodified RNA having the same sequence.
  • the effectiveness of the compositions and methods of the present disclosure are dependent on the presence of complement, while in another embodiment, the compositions and methods of the present disclosure are not dependent on the presence of complement.
  • the effectiveness of some of the compositions for use in the methods of the present disclosure are dependent on the presence of complement, while others are not.
  • the anti-gC antibody is dependent on complement for its effectiveness against HSV. 308 11979815v1 P-627574-PC [0511]
  • complement is an important contributor to innate and acquired immunity.
  • complement activation facilitates virus neutralization by particle phagocytosis and lysis, functions as a chemoattractant for neutrophils and macrophages, and enhances B and T cell responses.
  • HSV-1 gC, or an immunogenic fragment thereof binds complement C3b and blocks C5 and properdin interaction with C3b, which inhibit complement activation and complement-mediated virus neutralization.
  • a gC1 domain, or an immunogenic fragment thereof, that interacts with complement is located within amino acids 33 to 133 and blocks C5 and properdin binding to C3b, and in some embodiments, a gC1, or an immunogenic fragment thereof, domain that interacts with complement extends from amino acids 124 to 366 and directly binds C3b. In some embodiments, an HSV-1 gC mutant virus deleted in the C3b binding domain is more susceptible to complement-mediated virus neutralization in vitro and less virulent than wild-type (WT) virus in the mouse flank model.
  • WT wild-type
  • the interaction between gC1 and C3b enhances HSV-1 virulence, and in some embodiments, blocking the gC1 domain is effective in preventing or treating HSV-1 infection.
  • the compositions and methods of the present disclosure are for use in human subjects, while in another embodiment, they are for use in animal subjects.
  • the subject is mammalian.
  • the subject is any organism susceptible to infection by HSV.
  • the subject is murine, bovine, ovine, canine, feline, equine, porcine, etc.
  • the compositions and methods of the present disclosure are effective in male subjects.
  • the compositions and methods of the present disclosure are effective in female subjects.
  • compositions and methods of the present disclosure are effective in seronegative subjects. In another embodiment, the compositions and methods of the present disclosure are effective in seropositive subjects.
  • Pharmaceutical Formulations [0513] a method of present disclosure further comprises mixing an RNA with a transfection reagent prior to the step of contacting. In another embodiment, a method of present disclosure further comprises administering an RNA together with a transfection reagent. In another embodiment, the transfection reagent is a cationic lipid reagent.
  • the RNA can be one or more RNAs comprising a nucleotide sequence at least 80%, at least 85%, at least 309 11979815v1 P-627574-PC 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6.
  • a transfection reagent is a lipid-based transfection reagent.
  • a transfection reagent is a protein-based transfection reagent.
  • a transfection reagent is a polyethyleneimine based transfection reagent.
  • a transfection reagent is calcium phosphate.
  • a transfection reagent is Lipofectin® or Lipofectamine®.
  • a transfection reagent is any other transfection reagent known in the art.
  • a transfection reagent forms a liposome. Liposomes, in another embodiment, increase intracellular stability, increase uptake efficiency and improve biological activity.
  • liposomes are hollow spherical vesicles composed of lipids arranged in a similar fashion as those lipids which make up the cell membrane.
  • liposomes can deliver RNA to cells in a biologically active form (see Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; see generally ibid). [0517] Each type of transfection reagent represents a separate embodiment of the present disclosure.
  • an RNA of the present disclosure is encapsulated in a nanoparticle.
  • Methods for nanoparticle packaging are well known in the art, and are described, for example, in Bose S, et al., (Role of Nucleolin in Human Parainfluenza Virus Type 3 Infection of Human Lung Epithelial Cells. J. Virol. 78:8146. 2004); Dong Y et al., Poly(d,l-lactide-co- glycolide)/montmorillonite nanoparticles for oral delivery of anticancer drugs. Biomaterials 26:6068. 2005); Lobenberg R.
  • ⁇ RNA is encapsulated in nanoparticles to improve efficiency of delivery and expression of ⁇ RNA.
  • Nanoparticle packaging involves condensing and encapsulating RNA into particles that are smaller than the pore of the nuclear membrane, using chemicals including poly-L-lysine and polyethylene glycol.
  • RNA is packaged into one of four nanoparticle formulations (PEI, PLL, PAE, and CK 30 PEG 10k ).
  • nanoparticles used in the compositions and methods of the present disclosure comprise lipid nanoparticles as described in Cullis, P., & Hope, M. (n.d.). Lipid Nanoparticle Systems for Enabling Gene Therapies. Molecular therapy, 25(7), which is incorporated by reference herein in its entirety.
  • delivery of RNA comprises any suitable delivery method, including exemplary RNA transfection methods described elsewhere herein.
  • delivery of a nucleoside-modified RNA to a subject comprises mixing the nucleoside-modified RNA with a transfection reagent prior to the step of contacting.
  • a method of present disclosure further comprises administering nucleoside-modified RNA together with the transfection reagent.
  • the transfection reagent is a cationic lipid reagent.
  • RNAs can be one or more RNAs comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6.
  • the transfection reagent is a lipid-based transfection reagent.
  • the transfection reagent is a protein-based transfection reagent.
  • the transfection reagent is a polyethyleneimine based transfection reagent.
  • the transfection reagent is calcium phosphate.
  • the transfection reagent is Lipofectin®, Lipofectamine®, or TransIT®. In another embodiment, the transfection reagent is any other transfection reagent known in the art. [0523] In another embodiment, a transfection reagent forms a liposome. [0524] Liposomes, in another embodiment, increase intracellular stability, increase uptake efficiency and improve biological activity. In another embodiment, liposomes are hollow spherical vesicles composed of lipids arranged in a similar fashion as those lipids which make up the cell 311 11979815v1 P-627574-PC membrane.
  • the composition comprises a lipid nanoparticle (LNP) and one or more nucleic acid molecules described herein.
  • the composition comprises an LNP and one or more nucleoside-modified RNA molecules encoding one or more antigens, adjuvants, or a combination thereof.
  • lipid nanoparticle refers to a particle having at least one dimension on the order of nanometers (e.g., 1-1,000 nm) which includes one or more lipids, for example a lipid of Formula (I), (II) or (III), as described in WO2016176330A1, which is incorporated by reference herein in its entirety.
  • lipid nanoparticles are included in a formulation comprising a nucleoside-modified RNA as described herein.
  • such lipid nanoparticles comprise a cationic lipid and one or more excipient selected from neutral lipids, charged lipids, steroids and polymer conjugated lipids (e.g., a pegylated lipid such as a pegylated lipid of structure (IV), such as compound IVa).
  • the nucleoside-modified RNA is encapsulated in the lipid portion of the lipid nanoparticle or an aqueous space enveloped by some or all of the lipid portion of the lipid nanoparticle, thereby protecting it from enzymatic degradation or other undesirable effects induced by the mechanisms of the host organism or cells e.g. an adverse immune response.
  • the lipid nanoparticles have a mean diameter of from about 30 nm to about 150 nm, from about 40 nm to about 150 nm, from about 50 nm to about 150 nm, from about 60 nm to about 130 nm, from about 70 nm to about 110 nm, from about 70 nm to about 100 nm, from about 80 nm to about 100 nm, from about 90 nm to about 100 nm, from about 70 to about 90 nm, from about 80 nm to about 90 nm, from about 70 nm to about 80 nm, or about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm
  • the nucleoside-modified RNA when present in the lipid nanoparticles, is resistant in aqueous solution to degradation with a nuclease.
  • the LNP may comprise any lipid capable of forming a particle to which the one or more nucleic acid molecules are attached, or in which the one or more nucleic acid molecules are 312 11979815v1 P-627574-PC encapsulated.
  • lipid refers to a group of organic compounds that are derivatives of fatty acids (e.g., esters) and are generally characterized by being insoluble in water but soluble in many organic solvents.
  • Lipids are usually divided in at least three classes: (1) "simple lipids” which include fats and oils as well as waxes; (2) "compound lipids” which include phospholipids and glycolipids; and (3) "derived lipids” such as steroids.
  • the LNP comprises one or more cationic lipids, and one or more stabilizing lipids. Stabilizing lipids include neutral lipids and pegylated lipids.
  • the LNP comprises a cationic lipid.
  • the term "cationic lipid” refers to a lipid that is cationic or becomes cationic (protonated) as the pH is lowered below the pK of the ionizable group of the lipid, but is progressively more neutral at higher pH values. At pH values below the pK, the lipid is then able to associate with negatively charged nucleic acids.
  • the cationic lipid comprises a zwitterionic lipid that assumes a positive charge on pH decrease.
  • the cationic lipid comprises any of a number of lipid species which carry a net positive charge at a selective pH, such as physiological pH.
  • Such lipids include, but are not limited to, N,N-dioleyl-N,N-dimethylammonium chloride (DODAC); N-(2,3- dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA); N,N-distearyl-N,N- dimethylammonium bromide (DDAB); N-(2,3- dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP); 3-(N— ( ⁇ ', ⁇ '- dimethylaminoethane)-carbamoyl)cholesterol (DC-Choi), N- (l-(2,3-dioleoyloxy)propyl)- N-2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoracetate (DOSPA), dioctadecylamidoglycyl carboxy spermine (DO
  • cationic lipids are available which can be used in the present disclosure. These include, for example, LIPOFECTIN® (commercially available cationic liposomes comprising DOTMA and l,2-dioleoyl-sn-3- phosphoethanolamine (DOPE), from GIBCO/BRL, Grand Island, N.Y.); LIPOFECTAMINE® (commercially available cationic liposomes comprising N-(l-(2,3- dioleyloxy)propyl)-N-(2- (sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoroacetate (DOSPA) and (DOPE), from GIBCO/BRL); and TRANSFECTAM® (commercially available cationic lipids comprising dioctadecylamidoglycyl carboxyspermine (DOGS) in ethanol from Promega Corp., Madison, Wis.).
  • LIPOFECTIN® commercially available cationic liposome
  • the following lipids are cationic and have a positive charge at below physiological pH: 313 11979815v1 P-627574-PC DODAP, DODMA, DMDMA, l,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1 ,2- dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA).
  • the cationic lipid is an amino lipid. Suitable amino lipids useful in the disclosure include those described in WO 2012/016184, incorporated herein by reference in its entirety.
  • Representative amino lipids include, but are not limited to, l,2-dilinoleyoxy-3- (dimethylamino)acetoxypropane (DLin-DAC), l,2-dilinoleyoxy-3- morpholinopropane (DLin- MA), l,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), l,2-dilinoleylthio-3- dimethylaminopropane (DLin-S-DMA), l-linoleoyl-2-linoleyloxy-3- dimethylaminopropane (DLin-2-DMAP), l,2-dilinoleyloxy-3-trimethylaminopropane chloride salt (DLin-TMA.Cl), l,2- dilinoleoyl-3-trimethylaminopropane chloride salt (DLin-TAP.Cl), l,2-dilinoleyloxy-3-(N- methyl
  • the cationic lipid is present in the LNP in an amount from about 30 to about 95 mole percent. In some embodiments, the cationic lipid is present in the LNP in an amount from about 30 to about 70 mole percent. In some embodiments, the cationic lipid is present in the LNP in an amount from about 40 to about 60 mole percent. In some embodiments, the cationic lipid is present in the LNP in an amount of about 50 mole percent. In some embodiments, the LNP comprises only cationic lipids. In certain embodiments, the LNP comprises one or more additional lipids which stabilize the formation of particles during their formation. [0535] Suitable stabilizing lipids include neutral lipids and anionic lipids.
  • neutral lipid refers to any one of a number of lipid species that exist in either an uncharged or neutral zwitterionic form at physiological pH.
  • Representative neutral lipids include diacylphosphatidylcholines, diacylphosphatidylethanolamines, ceramides, sphingomyelins, dihydro sphingomyelins, cephalins, and cerebrosides.
  • Exemplary neutral lipids include, for example, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl- phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE) and dioleoyl-phosphatidylethanolamine 4-(N- 314 11979815v1 P-627574-PC maleimidomethyl)-cyclohexane-l- carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), di
  • the neutral lipid is l,2-distearoyl-sn- glycero-3- phosphocholine (DSPC).
  • the LNPs comprise a neutral lipid selected from DSPC, DPPC, DMPC, DOPC, POPC, DOPE and SM.
  • the molar ratio of the cationic lipid (e.g., lipid of Formula (I)) to the neutral lipid ranges from about 2 : 1 to about 8: 1.
  • the LNPs further comprise a steroid or steroid analogue.
  • the steroid or steroid analogue is cholesterol.
  • the molar ratio of the cationic lipid (e.g., lipid of Formula (I)) to cholesterol ranges from about 2:1 to 1:1.
  • anionic lipid refers to any lipid that is negatively charged at physiological pH.
  • lipids include phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N- dodecanoylphosphatidylethanolamines, N- succinylphosphatidylethanolamines, N- glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids.
  • the LNP comprises glycolipids (e.g., monosialoganglioside GMi).
  • the LNP comprises a sterol, such as cholesterol.
  • the LNPs comprise a polymer conjugated lipid.
  • polymer conjugated lipid refers to a molecule comprising both a lipid portion and a polymer portion.
  • An example of a polymer conjugated lipid is a pegylated lipid.
  • pegylated lipid refers to a molecule comprising both a lipid portion and a polyethylene glycol portion. Pegylated lipids are known in the art and include l-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG- s- DMG) and the like.
  • the LNP comprises an additional, stabilizing - lipid which is a polyethylene glycol-lipid (pegylated lipid).
  • Suitable polyethylene glycol- lipids include PEG- modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramides (e.g., PEG-CerC14 or PEG-CerC20), PEG-modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols.
  • Representative polyethylene glycol-lipids include PEG-c-DOMG, PEG-c-DMA, and PEG- s-DMG.
  • the polyethylene glycol-lipid is N-[(methoxy poly(ethylene glycol) 2 ooo)carbamyl]-l,2-dimyristyloxlpropyl-3-amine (PEG-c-DMA).
  • the polyethylene glycol-lipid is PEG-c-DOMG).
  • the LNPs comprise a pegylated diacylglycerol (PEG-DAG) such as l-(monomethoxy-polyethyleneglycol)-2,3- dimyristoylglycerol (PEG-DMG), a pegylated phosphatidylethanoloamine (PEG-PE), a PEG succinate diacylglycerol (PEG-S-DAG) such as 4-0-(2',3'-di(tetradecanoyloxy)propyl-l-0-(co- methoxy(polyethoxy)ethyl)butanedioate (PEG-S-DMG), a pegylated ceramide (PEG- cer), or a PEG dialkoxypropylcarbamate such as Q-methoxy(polyethoxy)ethyl-N-(2,3- di(tetradecanoxy)propyl)carbamate or 2,3-di
  • the molar ratio of the cationic lipid to the pegylated lipid ranges from about 100: 1 to about 25: 1.
  • the additional lipid is present in the LNP in an amount from about 1 to about 10 mole percent. In some embodiments, the additional lipid is present in the LNP in an amount from about 1 to about 5 mole percent. In some embodiments, the additional lipid is present in the LNP in about 1 mole percent or about 1.5 mole percent.
  • the LNP comprises one or more targeting moieties which are capable of targeting the LNP to a cell or cell population.
  • the targeting moiety is a ligand which directs the LNP to a receptor found on a cell surface.
  • the LNP comprises one or more internalization domains.
  • the LNP comprises one or more domains which bind to a cell to induce the internalization of the LNP.
  • the one or more internalization domains bind to a receptor found on a cell surface to induce receptor-mediated uptake of the LNP.
  • the LNP is capable of binding a biomolecule in vivo, where the LNP-bound biomolecule can then be recognized by a cell-surface receptor to induce internalization.
  • the LNP binds systemic ApoE, which leads to the uptake of the LNP and associated cargo.
  • systemic ApoE systemic ApoE
  • Other exemplary LNPs and their manufacture are described in the art, for example in WO2016176330A1, U.S. Patent Application Publication No.
  • methods of the present disclosure comprise administering an RNAs encoding one or more HSV glycoproteins, or immunogenic fragments thereof and a pharmaceutically acceptable carrier or diluent.
  • pharmaceutically acceptable carriers for liquid formulations may be aqueous or non-aqueous solutions, suspensions, emulsions or oils.
  • non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate.
  • Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media.
  • oils are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, and fish-liver oil.
  • pharmaceutically acceptable carriers or diluents are well known to those skilled in the art.
  • the pharmaceutical compositions provided herein are controlled- release compositions, i.e. compositions in which the compound is released over a period of time after administration.
  • Controlled- or sustained-release compositions include formulation in lipophilic depots (e.g., fatty acids, waxes, oils).
  • the composition is an immediate-release composition, i.e. a composition in which the entire compound is released immediately after administration.
  • each of the additives, excipients, formulations and methods of administration represents a separate embodiment of the present disclosure.
  • the present disclosure provides a kit comprising a reagent utilized in performing a method of the present disclosure.
  • the present disclosure provides a kit comprising a composition, tool, or instrument of the present disclosure. Certain Embodiments 1.
  • a combination comprising: (a) a first composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and 317 11979815v1 P-627574-PC (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, for use in a method of inhibiting, suppressing, or reducing the incidence of an HSV oral mucosal infection, an HSV encephalitis infection, or any combination thereof, in a subject.
  • HSV Herpes Simplex Virus
  • gC Herpes Simplex Virus
  • gE HSV glycoprotein E
  • a combination comprising: (a) a first composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, for use in a method of treating an HSV oral mucosal infection, an HSV encephalitis infection, or any combination thereof, in a subject.
  • HSV Herpes Simplex Virus
  • gC Herpes Simplex Virus
  • gE HSV glycoprotein E
  • a combination comprising: (a) a first composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, for use in a method of preventing establishment of a latent HSV infection in a subject.
  • HSV Herpes Simplex Virus
  • a combination comprising: (a) a first composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, (a) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (b) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, 318 11979815v1 P-627574-PC for use in a method of decreasing, suppressing, or inhibiting an HSV infection of the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or any combination thereof in a subject. 6.
  • HSV Herpes Simplex Virus
  • RNAs comprise one or more ribonucleic acids that are nucleoside modified.
  • nucleoside modified ribonucleic acids comprise one or more m 5 D (5-methyldihydrouridine) residues.
  • nucleoside modified ribonucleic acids comprise one or more pseudouridine residues.
  • said one or more pseudouridine residues comprise m1 ⁇ (1-methylpseudouridine).
  • any one of embodiments 1-10 wherein the HSV gC or an immunogenic fragment thereof is an HSV-2 gC or immunogenic fragment thereof, the HSV gD or an immunogenic fragment thereof is an HSV-2 gD or immunogenic fragment thereof, the HSV gE or an immunogenic fragment thereof is an HSV-2 gE or immunogenic fragment thereof, or any combination thereof. 12.
  • the HSV-2 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 28, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 241, or SEQ ID NO: 242;
  • the HSV-2 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 110, SEQ ID NO: 234, or SEQ ID NO: 120;
  • the HSV-2 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 17 or SEQ ID NO: 18.
  • the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 25, SEQ ID NO: 119, SEQ ID 319 11979815v1 P-627574-PC NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239, or SEQ ID NO: 246; the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 25, SEQ ID NO:
  • the HSV-2 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 224; the HSV-2 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 5; and/or the HSV-2 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 17.
  • RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 246; the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 247; and/or the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 145. 16.
  • RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 246;
  • the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 247;
  • the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 145. 320 11979815v1 P-627574-PC 17.
  • any one of embodiments 1-10 wherein the HSV gC or an immunogenic fragment thereof is an HSV-1 gC or immunogenic fragment thereof, the HSV gD or an immunogenic fragment thereof is an HSV-1 gD or immunogenic fragment thereof, and/or the HSV gE or an immunogenic fragment thereof is an HSV-1 gE or immunogenic fragment thereof. 18.
  • the HSV-1 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 240; the HSV-1 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 134; and/or the HSV-1 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 248, or SEQ ID NO: 140. 19.
  • RNA encoding HSV-1 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 24; the RNA encoding HSV-1 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 22; and/or the RNA encoding HSV-1 gE or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 26. 20.
  • the immunogenic fragment of HSV gC comprises the ectodomain of HSV gC
  • the immunogenic fragment of HSV gD comprises the ectodomain of HSV gD
  • the immunogenic fragment of HSV gE comprises the ectodomain of HSV gE. 21.
  • composition further comprises one or more RNAs encoding a) HSV glycoprotein B (gB) or immunogenic fragment thereof, b) HSV glycoprotein H (gH) or immunogenic fragment thereof, c) HSV glycoprotein L (gL) or immunogenic fragment thereof, d) HSV glycoprotein I (gI) or immunogenic fragment thereof, or e) any combination thereof.
  • RNAs encoding a) HSV glycoprotein B (gB) or immunogenic fragment thereof, b) HSV glycoprotein H (gH) or immunogenic fragment thereof, c) HSV glycoprotein L (gL) or immunogenic fragment thereof, d) HSV glycoprotein I (gI) or immunogenic fragment thereof, or e) any combination thereof. 22.
  • RNAs further comprise a signal sequence encoding a signal peptide. 321 11979815v1 P-627574-PC 23.
  • the signal sequence is or comprises any signal sequence listed in Table 4. 24.
  • 26. The combination of embodiment 24, wherein the viral signal peptide is an HSV signal peptide.
  • 27. The combination of embodiment 26, wherein the HSV signal peptide is an HSV gD signal peptide.
  • 28. The combination of embodiment 27, wherein the HSV gD signal peptide is an HSV-1 gD signal peptide. 29.
  • the combination of embodiment 28, wherein the HSV-1 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 34 or SEQ ID NO: 35. 30.
  • the combination of embodiment 28 or 29, wherein the HSV-1 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 35.
  • the combination of embodiment 31, wherein the HSV-2 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, or SEQ ID NO: 33.
  • the combination of embodiment 31 or 32, wherein the HSV-2 gD signal peptides comprises an amino acid sequence at least 85% identical to SEQ ID NO: 29 or SEQ ID NO: 31. 34.
  • 35. The combination of any one of embodiments 1-34, wherein one or more of said RNAs further comprise a poly-A tail.
  • 36. The combination of any one of embodiments 1-35, wherein one or more of said RNAs further comprise a cap.
  • 37. The combination of embodiment 36, wherein the cap is an m7GpppG cap, 3′-O-methyl- m7GpppG cap, or anti-reverse cap analog.
  • 38. The combination of any one of embodiments 1-35, wherein one or more of said RNAs further comprise a cap-independent translational enhancer. 322 11979815v1 P-627574-PC 39.
  • RNAs further comprise 5′ and/or 3′ untranslated regions.
  • 40. The combination of any one of embodiments 1-39, wherein one or more of said RNAs are encapsulated in or formulated with a nanoparticle, lipid, polymer, cholesterol, and/or cell penetrating peptide. 41. The combination of embodiment 40, wherein said nanoparticle is a liposomal nanoparticle. 42. The combination of any one of embodiments 1-41, wherein said HSV infection comprises an HSV-1 infection. 43. The combination of any one of embodiments 1-41, wherein said HSV infection comprises an HSV-2 infection. 44.
  • any one of embodiments 1-44 wherein one or more of said compositions is formulated for topical administration.
  • 51. The combination of any one of the embodiments 1-50, wherein each of the RNAs encoding the HSV glycoproteins are contained in a single composition. 323 11979815v1 P-627574-PC 52.
  • any one of embodiments 1-50, wherein the first composition, second composition, and third composition are the same composition.
  • 53. The combination of any one of embodiments 1-52, further comprising additional administrations of said compositions to be provided to a subject subsequent to the first administration.
  • 54. The combination of any one of embodiments 1-52, further comprising one or more doses of each of the first composition, second composition, or third composition.
  • 55. The combination of any one of embodiments 1-54, wherein said combination further comprises: (a) a first composition comprising HSV gD or an immunogenic fragment thereof, (b) a second composition comprising HSV gC or an immunogenic fragment thereof, and (c) a third composition comprising HSV gE or an immunogenic fragment thereof. 56.
  • HSV Herpes Simplex Virus
  • the combination comprises: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions. 59.
  • HSV Herpes Simplex Virus
  • a combination for the manufacture of a medicament for treating or inhibiting Herpes Simplex Virus (HSV) oral mucosal infection, an HSV encephalitis infection, or any combination thereof; or an HSV-1 infection wherein the combination comprises: 324 11979815v1 P-627574-PC (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions.
  • HSV Herpes Simplex Virus
  • embodiments 58 or 59 wherein the infection is a latent infection, a primary infection, or a secondary infection.
  • 61 The use of any one of embodiments 58-60, wherein the infection is in the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or any combination thereof. 62.
  • a combination for preventing establishment of a latent Herpes Simplex Virus (HSV) infection in a subject comprising: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions.
  • HSV Herpes Simplex Virus
  • a combination for the manufacture of a medicament for preventing establishment of a latent Herpes Simplex Virus (HSV) infection in a subject wherein the combination comprises: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, 325 11979815v1 P-627574-PC wherein the use comprises administering one or more doses of one or more of the compositions.
  • gD Herpes Simplex Virus
  • a combination for treating or inhibiting a Herpes Simplex Virus (HSV) infection of the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or a combination thereof in a subject wherein the combination comprises: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions.
  • HSV Herpes Simplex Virus
  • a combination for the manufacture of a medicament for treating or inhibiting a Herpes Simplex Virus (HSV) infection of the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or a combination thereof in a subject wherein the combination comprises: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions.
  • HSV Herpes Simplex Virus
  • Exemplary modified RNA (encoding an immunogenic gC2 fragment (SEQ ID NO: 326 11979815v1 P-627574-PC 10), encoding an immunogenic gD2 fragment (SEQ ID NO: 4), and encoding an immunogenic gE2 fragment (SEQ ID NO: 16)) was prepared based on the DNA coding sequences that encode HSV-2 glycoprotein C (gC2; amino acids 27-426 from HSV-2 strain 333; SEQ ID NO: 11), glycoprotein D (gD2; amino acids 26-331 from HSV-2 strain 333; SEQ ID NO: 5), and glycoprotein E (gE2; amino acids 24-405 from HSV-2 strain 2.12; SEQ ID NO: 17).
  • HSV-2 glycoprotein C gC2; amino acids 27-426 from HSV-2 strain 333; SEQ ID NO: 11
  • glycoprotein D gD2; amino acids 26-331 from HSV-2 strain 333; SEQ ID NO: 5
  • glycoprotein E gE2; amino acids 24-4
  • the exemplary modified RNA was incorporated into lipid nanoparticles (LNP) to prepare the following immunogens: (a) Poly C RNA in LNP; (b) a modified RNA encoding an exemplary immunogenic gC2 fragment in LNP; (c) a modified RNA encoding an exemplary immunogenic gD2 fragment in LNP; (d) a modified RNA encoding an exemplary immunogenic gE2 fragment in LNP; (e) modified RNA encoding exemplary immunogenic gC2, gD2, and gE2 fragments in LNP.
  • LNP lipid nanoparticles
  • Immunization groups were as follows: a) Controls (Poly C group): 10 ⁇ g Poly C RNA/LNP divided into 4 aliquots and administered at 4 separate sites. b) Exemplary gD2 immunogenic fragment alone (gD2 group): 10 ⁇ g RNA encoding an exemplary immunogenic gD2 fragment /LNP divided into 4 aliquots and administered at 4 separate sites.
  • mice were treated subcutaneously with 2mg Depo-Provera and 5 days later infected intravaginally with 5x10 3 PFU HSV-2 strain MS ( ⁇ 400 LD 50 ). On days 2 and 4 post-challenge, vaginal swabs were obtained for virus cultures.
  • mice in each vaccine group were sacrificed and dorsal root ganglia (DRG) excised for HSV-2 DNA qPCR. The remaining animals were evaluated for weight loss and hind limb weakness for 10 days while survival and genital disease were monitored for 28 days.
  • DRG dorsal root ganglia
  • EXAMPLE 2 CHARACTERIZATION OF TRANSLATIONAL PRODUCTS PRODUCED BY MODIFIED RNA ENCODING gC2, gD2, AND gE2 IMMUNOGENIC FRAGMENTS [0561] The ability of modified RNA to express proteins of the expected molecular weight when transfected into mammalian cells was verified.0.1 ⁇ g of modified RNA encoding exemplary gC2, gD2, or gE2 immunogenic fragments was transfected into 293T cells using TransIT-mRNA (Mirus Bio LLC) for the transfection. Eighteen hours later, cells were harvested and extracts prepared for Western blots.
  • TransIT-mRNA TransIT-mRNA
  • RNAs were designed to express immunogenic fragments of exemplary gC2, gD2 and gE2 ectodomain (labeled mRNA-ecto).
  • mRNA-ecto immunogenic fragments of exemplary gC2, gD2 and gE2 ectodomain
  • purified baculovirus proteins gC2, gD2, and gE2 expressing the same amino acids as the RNA constructs (labeled Bac-ecto) were used ( Figures 1A-C).
  • Immunization groups were as follows: Poly C (10 ⁇ g Poly C RNA/LNP divided into 4 aliquots and administered at 4 separate sites) (control); gD2 (10 ⁇ g RNA encoding exemplary gD2 immunogenic fragment/LNP divided into 4 aliquots and administered at 4 separate sites); Trivalent-I (10 ⁇ g RNA encoding exemplary gC2 immunogenic fragment /LNP, 10 ⁇ g RNA encoding exemplary gD2 immunogenic fragment /LNP, 10 ⁇ g RNA encoding exemplary gE2 328 11979815v1 P-627574-PC immunogenic fragment /LNP each divided into 2 aliquots and each given at 2 sites); and Trivalent-C (10 ⁇ g RNA encoding exemplary gC2 immunogenic fragment & 10 ⁇ g RNA encoding exemplary gD2 immunogenic fragment & 10 ⁇ g RNA encoding exemplary gE2 immunogenic fragment combined into LNP and divided into 4 aliquots and given at 4 sites).
  • the second immunization significantly boosted the ELISA titers compared to the first.
  • the differences between the titers in the second vs the first immunizations were significant, p ⁇ 0.05 (t-tests, comparing the antibody titers after the first and second immunization).
  • the RNA vaccine encoding gD2 and the trivalent RNA vaccines induced extremely high titers of ELISA antibodies after the first immunization that were significantly boosted after the second immunization.
  • EXAMPLE 4 BALANCED TH1 AND TH2 IgG ISOTYPES PRODUCED BY MODIFIED RNA IMMUNIZATION
  • T H 2 T H 2
  • IgG1 T H 2
  • IgG2a T H 1 antibodies
  • ELISA was performed on plates coated with all three antigens, gC2, gD2 and gE2. Serum obtained after the first or second immunization was added to the antigen-coated plates, and IgG1 or IgG2a was detected using HRP anti-mouse IgG1 or IgG2a.
  • IgG1 ( Figure 3A) and IgG2a ( Figure 3B) titers were significantly elevated after immunization with the gD2 vaccine and the trivalent modified RNA vaccines. Further, the IgG1 ( Figure 3A) or IgG2a ( Figure 3B) titers were significantly higher after the second modified RNA immunization compared to the first, p ⁇ 0.05 (t tests). [0567] Conclusion: The results demonstrate high titers of antibodies are produced to both IgG1 and IgG2a isotypes, indicating a balanced T H 1 and T H 2 response to immunization with modified RNA encoding exemplary gC2, gD2 and gE2 immunogenic fragments.
  • RNA groups While each of the RNA groups was not significantly different from one another, the trivalent vaccine given as a combined immunogen (Trivalent-C) performed the best of the three RNA groups ( Figure 4). [0569] Conclusion: Each of the modified RNA groups produced extremely high titers of neutralizing antibodies in the presence of 10% human complement. EXAMPLE 6: CD4+ AND CD8+ T CELL RESPONSES IN SPLENOCYTES AFTER MODIFIED RNA IMMUNIZATION [0570] Five animals from the trivalent modified RNA group that were immunized with each exemplary glycoprotein immunogenic fragment RNA at a separate site (Trivalent-I group) were euthanized 14 days after the second immunization. Splenocytes were prepared for T cell assays.
  • CD4 + T cells The modified RNA-expressed exemplary gC2, gD2, and gE2 subunit immunogenic fragments each stimulated polyfunctional CD4 + T cell responses ( Figures 5A-5B). Splenocytes harvested from immunized subjects and then stimulated with subunit glycoprotein immunogenic fragments increased polyfunctional CD4 + T cell responses ( Figure 5A).
  • CD8 + T cells Only exemplary gE immunogenic peptide pool 2 stimulated a significant IFN ⁇ CD8 + T cell response ( Figures 6A-6B).
  • HSV-2 VAGINAL TITERS AFTER MODIFIED RNA IMMUNIZATION AND INTRAVAGINAL CHALLENGE Vaginal swabs were obtained from 10 animals per group on days 2 and 4 post challenge and cultured for replication competent HSV-2 virus. Results are shown in Figures 8A-8B.
  • EXAMPLE 10 HSV-2 DNA IN DORSAL ROOT GANGLIA AFTER MODIFIED RNA IMMUNIZATION AND INTRAVAGINAL CHALLENGE [0579] Five animals per group were euthanized at 4 days post challenge, except for the trivalent- combined group in which four animals were euthanized. Dorsal root ganglia (DRG) were harvested for HSV-2 DNA quantitation by qPCR to detect the Us9 gene.
  • DRG Dorsal root ganglia
  • the poly C RNA controls received 10 ⁇ g Poly C RNA-LNP divided into 4 aliquots and administered at 4 separate sites.
  • the trivalent modified RNA group received 10 ⁇ g RNA encoding exemplary gC2 immunogenic fragment/LNP, 10 ⁇ g modified RNA encoding exemplary gD2 immunogenic fragment/LNP, and 10 ⁇ g modified RNA encoding exemplary gE2 immunogenic fragment/LNP each divided into 2 aliquots and each given at 2 sites.
  • Two weeks after the second immunization spleens were harvested from 5 animals per group and flow cytometry performed to detect T follicular helper (Tfh) cells (Figure 11A; *p ⁇ 0.05) and germinal center B cell responses (Figure 11B; *p ⁇ 0.05).
  • EXAMPLE 12 VAGINAL IgG RESPONSES TO MODIFIED RNA IMMUNIZATION IN MICE [0584] BALB/c mice were immunized intradermally twice at 28 day intervals with 10 ⁇ g of poly C RNA-LNP, 10 ⁇ g modified RNA encoding exemplary gD2 immunogenic fragment/LNP or 10 ⁇ g trivalent modified RNA encoding each of exemplary gC2, gD2, and gE2 immunogenic fragments/LNP.
  • the trivalent modified RNA was combined and administered as 10 ⁇ g modified RNA encoding exemplary gC2 immunogenic fragment, 10 ⁇ g modified RNA encoding exemplary gD2 immunogenic fragment, and 10 ⁇ g modified RNA encoding exemplary gE2 immunogenic fragment combined into LNP and divided into 4 aliquots and given at 4 sites.
  • 60 ⁇ l of media were introduced in the vaginal cavity and retrieved.
  • the trivalent modified RNA produced a robust vaginal IgG response to gC2 ( Figure 12A) and gD2 ( Figure 12B) and a more moderate response to gE2 ( Figure 12C).
  • the gD2 ELISA titers were higher in mice immunized with the trivalent RNA vaccine compared to mice immunized with the gD2 RNA vaccine ( Figure 12B).
  • EXAMPLE 13 ANTIBODIES TO EXEMPLARY gC2 IMMUNOGENIC FRAGMENT PRODUCED BY TRIVALENT RNA IMMUNIZATION OF MICE BLOCK IMMUNE EVASION DOMAINS ON gC2
  • BALB/c mice were left unimmunized as a source of non-immune IgG, or immunized intradermally with poly C RNA/LNP or trivalent modified RNA/LNP.
  • the poly C RNA controls received 10 ⁇ g poly C RNA/LNP divided into 4 aliquots and administered at 4 separate sites.
  • the gD2 group received 10 ⁇ g modified RNA encoding exemplary gD2 immunogenic fragment/LNP administered as described for the poly C RNA/LNP.
  • the trivalent modified RNA group received 10 ⁇ g modified RNA encoding exemplary gC2 immunogenic fragment/LNP, 10 ⁇ g modified RNA encoding exemplary gD2 immunogenic fragment/LNP, and 10 ⁇ g modified RNA encoding exemplary gE2 immunogenic fragment/LNP combined into one LNP and divided into 4 aliquots and given at 4 sites. There were 10 mice in each group. Sera from the 10 mice were pooled and IgG was purified. The IgG was evaluated at 12 ⁇ g/200 ⁇ l for its ability to block complement component C3b binding to gC2.
  • This blocking assay is used to assess whether antibodies produced by immunization block the immune evasion properties of gC2.
  • Non-immune murine IgG, IgG from the poly C RNA group, and IgG from the gD2 group each failed to block gC2 binding to C3b.
  • IgG from trivalent modified RNA-immunized animals totally blocked the interaction between gC2 and C3b ( Figure 13, ****p ⁇ 0.0001).
  • the trivalent modified RNA vaccine produces antibodies that block immune evasion domains on gC2 as determined by blocking the interaction between gC2 and C3b.
  • EXAMPLE 14 INTRAVAGINAL INFECTION OF MICE AT A HIGHER INOCULUM TITER OF HSV-2 AFTER MODIFIED RNA VACCINATION
  • mice were treated with medroxyprogesterone and five days later infected intravaginally with 5x10 4 PFU HSV-2 strain MS (2,000 LD 50 ). Animals were followed for 28 days and evaluated for death, genital disease, vaginal viral titers 2 and 4 days after infection and dorsal root ganglia (DRG) HSV-2 DNA copy number 28 days after infection. No mouse immunized with the trivalent modified RNA/LNP vaccine died, had genital disease, had any virus detected on day 2 or 4 post-infection or had HSV-2 DNA detected in DRG (Table 8). Table 8.
  • mice Trivalent modified RNA/LNP immunized mice challenged with HSV-2 strain MS (2,000 LD50) Disease parameters Mice % Protection Death 0/5 100 [0589] Co c us o s: ce were n ec e w - a a ose a was - o gher than used in earlier experiments described herein ( Figures 7A-10). Protection of the mice remained outstanding even at this higher titer challenge. All five mice achieved sterilizing immunity as determined by no deaths, no genital disease, negative vaginal virus titers on days 2 and 4 post- infection and no HSV-2 DNA in the lumbosacral DRG on day 28 (Table 8).
  • EXAMPLE 15 EVALUATION OF THE INTRAMUSCULAR ROUTE OF MODIFIED RNA IMMUNIZATION IN MICE [0590]
  • BALB/c mice were immunized intramuscularly with poly C RNA/LNP as a control (15/group) or with trivalent modified RNA containing 10 ⁇ g of modified RNA encoding each of exemplary gC2, gD2, and gE2 immunogenic fragments/LNP (20/group). All poly C control animals died by day 12, while all animals in the trivalent modified RNA group survived (Figure 14A). No weight loss occurred in the trivalent modified RNA group, while the poly C control animals lost >15% of body weight ( Figure 14B).
  • the poly C group developed extensive genital disease, while the trivalent modified RNA animals had no genital disease (Figure 14C).
  • DRG were harvested from nine poly C animals at the time of euthanasia between days 7 and 12 post- infection or at the end of the experiment on day 28 in the trivalent modified RNA group. All 335 11979815v1 P-627574-PC animals in the poly C group had HSV-2 DNA detected in DRG, while none were positive for HSV- 2 DNA in trivalent modified RNA group (Figure 14D).
  • Day 2 ( Figure 14E) and Day 4 (Figure 14F) vaginal viral cultures were positive in all 15 animals in the poly C group, while cultures were negative in all 20 animals in the trivalent modified RNA group.
  • Trivalent modified RNA/LNP provides outstanding protection in mice when administered intramuscularly. Comparable findings to the above were observed when mice were immunized intradermally. Overall, 64 mice have been evaluated, which were immunized with trivalent modified RNA at 10 ⁇ g of each immunogenic fragment given either intradermally ( Figures 7A-10) or intramuscularly ( Figures 14A-14F). Sterilizing immunity was achieved in 63/64 (98%) mice based on no death, no genital disease, no weight loss, negative day 2 and 4 vaginal titers and negative HSV-2 DNA in DRG.
  • EXAMPLE 16 SUMMARY COMPARISON OF IMMUNIZATION WITH TRIVALENT RNA/LNP AND TRIVALENT SUBUNIT ANTIGEN CPG/ALUM IN BALB/c MICE [0592]
  • the results presented in Table 9 hereinbelow represent a summary of all the results in BALB/c mice that were immunized either intradermally or intramuscularly with trivalent modified RNA containing 10 ⁇ g of modified RNA encoding gC2, gD2 and gE2 immunogenic fragments/LNP (total 64 mice studied).
  • mice that were immunized with 5 ⁇ g each of bac-gC2(27-426t) containing gC2 amino acids 27-426 from HSV-2 strain 333, bac-gD2(306t) containing gD2 amino acids 26-331 from HSV-1 strain 333, and bac-gE2(24-405t) containing gE2 amino acids 24-405 from HSV-2 strain 2.12.
  • the exemplary gC2, gD2, gE2 fragment antigens were mixed with 150 ⁇ g CpG and 25 ⁇ g alum/per ⁇ g protein as adjuvants and administered intramuscularly.
  • mice were immunized twice at 28-day intervals with trivalent modified RNA/LNP and three times at 14-day intervals with exemplary gC2, gD2, gE2 fragment antigens, as done in prior experiments.
  • the modified RNA and gC2, gD2, gE2 fragment antigen experiments were performed at the same time.
  • Table 9 demonstrate significant superiority of the trivalent modified RNA/LNP vaccine over the trivalent gC2, gD2, gE2 fragment antigen vaccine in many immune response parameters, and most importantly in vaccine efficacy.
  • the trivalent modified RNA/LNP vaccine achieved sterilizing immunity in 63/64 (98%) of mice compared to 15/20 (75%) in the subunit antigen group.
  • EXAMPLE 17 EVALUATION OF THE TRIVALENT MODIFIED RNA/LNP VACCINE IN GUINEA PIGS
  • animals in the poly C and trivalent modified RNA groups were infected intravaginally with 5x10 5 PFU of HSV-2 strain MS (50 LD50).
  • EXAMPLE 18 EVALUATION OF THE HSV-1 AND HSV-2 TRIVALENT NUCLEOSIDE-MODIFIED RNA/LNP VACCINES AGAINST GENITAL AND NON- GENITAL HSV-1 INFECTION IN MICE [0595]
  • the present Example demonstrates that an RNA vaccine encoding exemplary HSV gC, gD, and gE immunogenic fragments is effective against genital HSV-1 and HSV-2 infection, as well as HSV-1 oral infection.
  • Tri-HSV-2 RNA vaccine encoding HSV-2 gC, gD, and gE
  • tri-HSV-1 RNA vaccine encoding HSV-1 gC, gD, and gE
  • the tri- HSV-2 immunogens were prepared to generate RNA that encodes exemplary immunogenic fragments of the ectodomains of HSV-2 gC (27-426; SEQ ID NO: 11), HSV-2 gD (26-333; SEQ ID NO: 120), and HSV-2 gE (24-405; SEQ ID NO: 17).
  • the tri-HSV-1 exemplary immunogens were prepared to generate RNA that encodes exemplary immunogenic fragments of the ectodomains of the HSV-1 gC (27-457; SEQ ID NO: 8), HSV-1 gD (26-333; SEQ ID NO: 134), and HSV-1 gE (24-309; SEQ ID NO: 140) proteins where the first amino acid listed represents the amino acid immediately after the signal sequence. Each subunit protein extends from the first amino acid after the signal sequence to just prior to the transmembrane domain.
  • TriLink m1 ⁇ -5′-triphosphate
  • RNA was capped using the m7G capping kit with 2′-O-methyltransferase (ScriptCap, CellScript).
  • the RNA contains a 101-nucleotide poly(A) tail.
  • Nucleoside-modified RNAs were purified by Fast Protein Liquid Chromatography (FPLC) (Akta Purifier GE Healthcare) and stored at -20°C.
  • FPLC nucleoside-modified RNAs and polyC RNA (Sigma) were encapsulated in LNPs using a self-assembly process in which an aqueous solution of RNA at acidic pH 4.0 was rapidly mixed with a solution of lipids dissolved in ethanol.
  • LNPs used in this study were similar in composition to those described previously, which contain an ionizable cationic lipid/phosphatidylcholine/cholesterol/PEG-lipid (Acuitas) (50:10:38.5:1.5 mol/mol) and were encapsulated at an RNA to total lipid ratio of ⁇ 0.05 (wt/wt).
  • the LNPs had a diameter of ⁇ 80 nm as measured by dynamic light scattering using a Zetasizer Nano ZS (Malvern Instruments Ltd, Malvern, UK) instrument, and were stored at -80°C at a concentration of RNA of ⁇ 1 ⁇ g/ ⁇ l.
  • mice Female BALB/c mice aged 6-8 weeks old were immunized twice 30 days apart in the hind right gastrocnemius muscle. Mice were immunized with either 1 ⁇ g or 10 ⁇ g (total) of tri-HSV-1 or tri-HSV-2 modified RNA vaccines. Control mice were injected with an equal volume of sterile saline.
  • Serum was collected 25 days after the final immunization and stored at -80 °C.
  • Serum IgG ELISA endpoint titers Purified baculovirus-derived exemplary gC1, gD1, and gE1 immunogenic fragments were produced as previously described. Purified protein was added 339 11979815v1 P-627574-PC to High Binding Costar microtiter plates. Serial 2-fold dilutions of serum starting at 1:250 were added to protein coated wells. Anti-mouse IgG was detected by adding 2,2’-azino-bis-3- ethylbenzothiazoline-6-sulfonic acid (ABTS).
  • ABTS 2,2’-azino-bis-3- ethylbenzothiazoline-6-sulfonic acid
  • Endpoint titers were calculated as the serum concentration that had an absorbance value of ⁇ 0.1 and was ⁇ two-fold higher than serum from PBS immunized mice.
  • HSV-1 and HSV-2 neutralizing antibody titers were measured by incubating 2-fold serial dilutions of mouse sera with 100 PFU of HSV-1 strain NS at 37°C for 1 hour. Incubations were done in the presence of 5% human serum as a source of complement obtained from an HSV-1/HSV-2 seronegative donor. Remaining infectious virus was determined by plaque assay on Vero cells. The neutralizing titer was defined as the highest serum dilution that reduced plaques by ⁇ 50%.
  • mice Isolation and stimulation of splenocytes from BALB/c mice. Mice were immunized twice with PBS or 10 ⁇ g of exemplary tri-HSV-1 or tri-HSV-2 vaccine. Immunized mice were sacrificed 10 days after the second immunization. The spleens were harvested, and mechanically dissociated into single-cell suspensions. Cell counts were obtained and 2 ⁇ 10 6 cells were added to individual wells for each stimulation condition for each mouse. Splenocytes were stimulated in RPMI media supplemented with 55 ⁇ M 2-mercaptoethanol and 10 units recombinant mouse IL-2/ ⁇ L stimulation media. Intracellular cytokines were immobilized with 5 ⁇ g/mL Brefeldin A and Monensin.
  • DMSO Dimethyl Sulfoxide
  • PMA phorbol-12- myristate 13-acetate
  • ionomycin 0.67 ⁇ M ionomycin
  • 1 ⁇ g/mL of HSV-1 protein overlapping peptide pools Cells were incubated for 12 hours at 37 °C in 5% CO 2 .
  • Overlapping peptide pools Overlapping peptide pools were synthesized by GenScript with a purity of ⁇ 70%. Peptides were 15 amino acids long with an 11 amino acid overlap. Identical amino acid sequences used in the modified RNA encoding exemplary immunogenic fragments were used for synthesizing overlapping peptide sequences.
  • Antibodies used for labeling cells were fluorescein isothiocyanate (FITC) conjugated anti-CD3 (clone 17A2), allophycocyanin (APC)-Fire 750 conjugated to CD4 (Clone GK1.5), phycoerythrin (PE) conjugated anti-IL-2 (cloneJES6-5H4), PE-Dazzle 594 conjugated anti-CD107a (clone 1D4B), Peridinin-Chlorophyll-Protein-Cyanin (PerCP-Cy5.5) conjugated anti-CD8 ⁇ (clone 53-6.7), Alexa-Flour (AF)-700 conjugated anti-IFN ⁇ (clone XMG1.2), Brilliant Violet 421 conjugated 340 11979815v1 P-627574-PC anti-IL-4 (Clone 11b11) and PE-Cy7 conjugated anti-TNF ⁇ (clone MP6-XT22).
  • FITC fluorescein isothiocyanate
  • Dead cells were discriminated by Fixable Live-Dead Aqua dye (Invitrogen). Brefeldin A, PMA/ionomycin, Cyto- fast-fix-perm buffer, and intracellular staining permeabilization wash buffer were purchased from BioLegend. After labeling of intracellular cytokines, cells were fixed with 1% formaldehyde in PBS and stored at 4°C until the next day. Cells were analyzed on a BD LSR 2. Gates were set to discriminate singlet cells and then lymphocytes based on forward scatter and side scatter properties. 150,000 events in the lymphocyte gate were collected. After data collection, flow samples were analyzed using FlowJo version 10.8 (Tree Star software).
  • HSV-1 lip infection Mice were challenged 30 days after the 2 nd immunization. Mice were anesthetized with a solution of 100 mg/kg Ketamine and 12.5 mg/kg Xylazine, placed on their back, and a 25G needle was used to make 10 vertical scratches on the lower lip. After scratching the lip, 2 ⁇ 10 6 PFU of HSV-1 strain NS was topically applied. Mice were monitored for 13 days for weight loss and presence of lesions on the lower lip. [0606] Lip and Trigeminal (TG) tissue isolation.
  • the lip and TG were removed and placed in tubes containing 500 ⁇ L DMEM supplemented with 5% FBS. The tissues were stored at -80 °C until further processing. Lip and TG tissues were subjected to 3 freeze-thaw cycles. The lip was homogenized in a glass grinder and resuspended in 1 mL of media. The TG was homogenized using the frosted ends of glass slides and resuspended in 275 ⁇ L of DMEM. Tissue homogenates were used to make 10-fold serial dilutions that were then evaluated by plaque assay on Vero cells. [0607] HSV-1 vaginal infection. Mice were challenged with 2 ⁇ 10 6 PFU of HSV-1 strain NS intravaginally.
  • mice were scored daily for 14 days on a basis of one point each for redness/erythema, hair loss, distended belly indicating loss of fecal motility, and urinary staining for a maximum daily score of four.
  • the vaginal canals of mice were swabbed on days 2 and 4 for virus titers.
  • Vaginal swabs were placed in 1 mL of DMEM supplemented with L-glutamine, HEPES, antibiotics, and 5% FBS. Serial 10-fold dilutions were evaluated by plaque assay on Vero cells. [0608] HSV-1 DNA isolation.
  • the DRG were harvested at the time of sacrifice of the mice either at the end of the experiment on day 28 post-challenge or prior to day 28 in mice in which humane euthanasia was required.
  • Purified DNA was isolated and amplified for qPCR as previously described and the HSV-1 DNA copy number was expressed as log10 DNA copies per 10 5 adipsin 341 11979815v1 P-627574-PC genes. Samples with less than one copy by 40 cycles in duplicate wells were considered negative. If only one of the duplicate wells was positive, the sample was tested in triplicate and considered positive if two or more of the triplicates were positive. [0609] Statistics. The log-rank test was used to calculate P values for survival curves.
  • the Mann- Whitney test was used to calculate P values in experiments evaluating vaginal titers, serum antibodies by ELISA, HSV-1 neutralizing titers, viral titers present in lip and TG tissue, and HSV- 1 DNA copy number in the lip, TG, and DRG.
  • the flow cytometry data was analyzed using 2-way ANOVA with correction for multiple comparisons. All significance tests were two-tailed with a P value ⁇ 0.05 considered significant. Analysis was done using GraphPad Prism version 9.5 (GraphPad software Inc). Results [0610] The efficacy of exemplary tri-HSV-1 vaccine and exemplary tri-HSV-2 vaccine against HSV-1 challenge to lip or vagina was compared.
  • mice were immunized with either a 1 ⁇ g (low) or 10 ⁇ g (high) dose of total modified RNA containing equal concentrations of each modified RNA immunogen.
  • the tri-HSV-1 immunized mice had significantly higher serum IgG ELISA antibodies to gC1 and gE1 than tri- HSV-2 immunized animals ( Figures 16A-16B).
  • the gC1 IgG levels were 4.6-fold higher in the tri-HSV-1 group than in the tri-HSV-2 group at 1 ⁇ g and 6.9-fold higher at 10 ⁇ g doses.
  • a dose response with the tri-HSV-1 vaccine was detected in that the gC1 IgG levels were 2.3-fold higher in animals immunized with 10 ⁇ g than 1 ⁇ g, while no differences were detected at the two doses in the tri-HSV-2 immunized mice ( Figure 16A).
  • the gE1 IgG levels were 35- and 72-fold higher in tri-HSV-1 than tri-HSV-2 immunized mice at the low and high doses, respectively.
  • a 2.5-fold dose-response increase in gE1 IgG titers in the high dose tri-HSV-1 immunized mice ( Figure 16B) was detected.
  • the gD1 IgG levels were not significantly different comparing tri-HSV-1 and tri- HSV-2 immunized mice at the low or high dose (Figure 17C).
  • the IgG ELISA results suggest that many epitopes on gD1 and gD2 are type common, while epitopes on gC1 and gC2, or gE1 and gE2 are type specific.
  • Sera was evaluated for HSV-1 neutralizing antibodies in the presence of 5% human serum as a source of complement. Neutralizing titers were higher in the tri-HSV-1 vaccine immunized mice at both immunization doses, but differences did not reach statistical significance (Figure 16D).
  • the neutralizing antibody results are consistent with the gD ELISA results showing comparable IgG binding titers to gD1 produced by the tri-HSV-1 and tri-HSV-2 vaccines.
  • the 342 11979815v1 P-627574-PC neutralizing titers are also consistent with the prior observation that gD antibodies are more potent at neutralizing virus than gC or gE antibodies.
  • Tri-HSV-2 immunization induces a more robust CD8+ T-cell responses to gE1 overlapping peptides than tri-HSV-1 immunization.
  • the T-cell response in mice immunized twice with 10 ⁇ g tri-HSV-1 or tri-HSV-2 vaccines was evaluated.
  • the number of IFN ⁇ + TNF ⁇ + (double positive) CD8 + T-cells was 3.4-fold higher in the tri-HSV-2 immunized mice than the tri-HSV-1 immunized vaccine group ( Figure 18B, 3 rd group from right). Additionally, the number of triple positive CD8+ T-cells (IFN ⁇ + TNF ⁇ + IL-2 + ) was 8.2-fold higher in the tri- HSV-2 vaccine immunized group compared to the tri-HSV-1 vaccine immunized group ( Figure 18B, last group on right). CD8+ T-cell activity was analyzed by CD107a labeling, a marker of degranulation.
  • the tri-HSV-1 vaccine immunized group had more activated T-cells compared to the PBS group, but the tri-HSV-2 vaccine immunized animals had 7-fold greater CD107a labeling compared to the tri-HSV-1 vaccine immunized animals (Figure 18C).
  • CD4+ T-cells stimulated with gD1 overlapping peptides were analyzed.
  • CD4+ T-cells in the tri-HSV-2 vaccine immunized group had a 2-fold increase in polyfunctional cells (IFN ⁇ + TNF ⁇ + IL-2 + ) compared to tri-HSV-1 vaccine immunized mice (Figure 18D).
  • the 10 ⁇ g tri-HSV-2 modified RNA vaccine immunized group had a significantly lower copy number of viral DNA compared to the 10 ⁇ g tri-HSV-1 modified RNA vaccine immunized group ( Figure 19B).
  • Infectious virus in the TG was measured 5 days post-challenge.
  • Five of 10 (50%) PBS mice had replication competent virus present in the TG, while no immunized mouse had replication competent virus ( Figure 19C).
  • the presence of replication competent virus or HSV-1 DNA in the TG indicates a risk of virus establishing a latent infection.
  • Four of 10 (40%) of the PBS mice had viral DNA in the TG at 5 days ( Figure 19D).
  • mice immunized with tri-HSV-2 modified RNA vaccine had positive day 2 vaginal virus titers (mean titer 1.3 PFU/mL) after HSV- 1 intravaginal challenge.
  • An evaluation of whether the tri-HSV-1 modified RNA vaccine was better than the tri-HSV-2 modified RNA vaccine in reducing day 2 or day 4 vaginal virus titers after HSV-1 challenge was performed.
  • mice were vaccinated with tri-HSV-1 and tri-HSV-2 modified RNA vaccines at low (1 ⁇ g) and high (10 ⁇ g) doses, treated with Depo-Provera five days before infection, and challenged with 2 ⁇ 10 6 PFU of HSV-1 strain NS.
  • the PBS immunized animals started succumbing to infection by day 8. By day 14, only 1 of 5 (20%) survived the HSV- 1 challenge, while none of the tri-HSV-1 or tri-HSV-2 modified RNA vaccinated mice died, at either1 ⁇ g or 10 ⁇ g doses (Figure 20A).
  • mice started to lose weight by 6 days post-challenge, while the tri-HSV-1 and tri-HSV-2 modified RNA vaccine immunized mice had little or no weight loss (Figure 20B).
  • Mice were scored for genital disease over 14 days.
  • the PBS vaccinated mice developed genital disease on day 5 post-challenge that peaked on day 11 with a mean disease score of 2.8. Genital disease was not observed in any vaccinated mouse (Figure 20C).
  • An evaluation of whether infectious virus was present in vaginal swabs obtained on days 2 or 4 post-challenge was performed.
  • the tri-HSV-2 RNA vaccine immunization provided total protection against clinical and subclinical infection. Importantly, tri-HSV-2 modified RNA vaccine immunized mice were completely protected against HSV-1 DNA reaching the TG, indicating a minimal risk of latency and reactivation. [0621] An assessment was performed to determine whether the tri-HSV-1 modified RNA vaccine provides better protection than the tri-HSV-2 modified RNA vaccine in the murine genital infection model, particularly against subclinical infection measured by day 2 and day 4 vaginal virus titers.
  • mice T-cells play a vital role in restricting the ability of HSV-1 or HSV-2 to reactivate from latency. Mice immunized with tri-HSV-2 modified RNA were shown to have robust CD8+ T-cell responses to gE1 overlapping peptides.
  • the responses generated by tri-HSV-2 modified RNA vaccine immunized mice were stronger than by tri-HSV-1 modified RNA vaccine immunized mice as manifest by an increase of polyfunctional IFN ⁇ + TNF ⁇ + IL-2 + CD8 + T-cells and an increase in CD107a + CD8 + T-cells.
  • a greater CD4+ T cell response to gD1 overlapping peptides was detected in tri-HSV-2 modified RNA vaccine immunized mice than tri-HSV-1 modified RNA vaccine immunized mice.
  • These T cell responses may translate into the tri-HSV-2 modified RNA vaccine having an advantage over the tri-HSV-1 modified RNA vaccine in preventing HSV-1 reactivation from latency.
  • This advantage may be more apparent in the guinea pig genital infection model than the mouse model because recurrent genital lesions and recurrent shedding of HSV DNA can be detected in the guinea pig model.
  • antibody responses to gC1 and gE1 were lower for the tri-HSV-2 modified RNA vaccine than the tri-HSV-1 modified RNA vaccine, the T- cell responses were more robust.
  • the potent CD8+ T cell responses reveal an unexpected benefit of the tri-HSV-2 modified RNA vaccine against HSV-1 infection.
  • HSV-1 is the most common cause of sporadic infectious encephalitis worldwide and carries approximately a 30% mortality and 60% to 70% morbidity in survivors despite treatment with acyclovir. A vaccine that prevents HSV-1 encephalitis would represent a major advance.
  • mice at age 6-8 weeks were immunized intramuscularly in a hind limb twice at one-month intervals with PBS (control), with 1 ⁇ g trivalent HSV-2 RNA vaccine containing 0.33 ⁇ g each of exemplary gC2, gD2, gE2 nucleoside-modified RNA in lipid nanoparticle (LNP), or with 10 ⁇ g trivalent HSV-2 RNA vaccine containing 3.33 ⁇ g each of exemplary gC2, gD2, gE2 nucleoside-modified RNA in LNP.
  • mice were infected intranasally with 5x10 5 PFU of HSV-1 (100 LD 50 ) and evaluated for survival, neurologic disease, and for HSV-1 DNA copy number by qPCR in brain, trigeminal ganglia, and olfactory bulb tissues harvested post-infection.
  • Na ⁇ ve (not vaccinated) 6 to 8-week-old female mice were inoculated intranasally with 5x10 3 , 5x10 4 , 5x10 5 , 5x10 6 plaque-forming units (pfu) of HSV-1 strain H129, or with PBS 347 11979815v1 P-627574-PC as a control. Survival was evaluated over 28 days.
  • mice in the PBS group survived, while all mice in the groups inoculated with 5x10 4 , 5x10 5 , or 5x10 6 pfu required humane euthanasia (Figure 21). Survival was 50% (2 of 4 mice) at 5x10 3 pfu, establishing the LD 50 as 5x10 3 pfu ( Figure 21).
  • Trivalent HSV-2 vaccine protects BALB/c mice against HSV-1 encephalitis. All 10 animals in the vaccine group immunized with 1 ⁇ g or 10 ⁇ g of the tri-HSV-2 RNA-LNP survived, while all 10 mice in the PBS group required humane euthanasia on day 6 post-infection ( Figure 22A).
  • HSV-1 DNA was detected in brains in 5 of 5 (100%) mice in the PBS group with a mean titer of 7 log 10 HSV-1 DNA copies per 10 5 adipsin genes, while HSV-1 DNA was detected in 2 of 10 (20%) mice in the 1 ⁇ g group with a mean titer of 0.4 log 10 , or 1 of 10 (10%) mice in the 10 ⁇ g group with a mean titer of 0.2 log 10 ( Figure 22B).
  • HSV-1 DNA was detected in the trigeminal ganglia in 5 of 5 (100%) mice from the PBS group at a mean titer of 6.7 log 10, while HSV-1 DNA was detected in 9 of 10 (90%) mice in the 1 ⁇ g group with a mean titer of 2.8 log 10 , or 7 of 10 (70%) mice in the 10 ⁇ g group with a mean titer of 2.0 log10 (Figure 22C).
  • HSV-1 DNA was detected in the olfactory bulb of mice in the PBS group in 5 of 5 (100%) mice at a mean titer of 7.2 log 10, while HSV-1 DNA was detected in 2 of 9 (22%) mice in the 1 ⁇ g group with a mean titer of 0.6 log 10 , or 2 of 10 (20%) mice in the 10 ⁇ g group with a mean titer of 0.4 log 10 ( Figure 22D).
  • the 1 and 10 ⁇ g doses of the trivalent HSV-2 vaccine were both highly efficacious at protecting mice against death and preventing HSV-1 DNA from reaching the brain and the olfactory bulb.
  • the vaccine was less effective at preventing HSV-1 DNA from reaching the trigeminal ganglia, although the vaccine groups had significantly reduced HSV-1 DNA copy number in the trigeminal ganglia compared with the PBS group.
  • the HSV-2 nucleoside-modified RNA-LNP vaccine is highly efficacious at preventing HSV-1 encephalitis in a mouse model following a high dose lethal challenge.

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Abstract

The present disclosure provides compositions for the prevention and treatment of genital herpes, comprising nucleoside modified RNAs that encode herpes simplex virus (HSV) glycoproteins, including those involved in virus entry and immune evasion, and methods of use thereof.

Description

P-627574-PC RNA VACCINES ENCODING HERPES SIMPLEX VIRUS GLYCOPROTEINS AND USES THEREOF SEQUENCE LISTING STATEMENT [0001] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. The XML copy, created on June 2, 2024, is named P-627574-PC-SQL-02JUN24.xml and is 379 kilobytes in size. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT [0002] This invention was made with U.S. government support under AI139618 awarded by The National Institutes of Health. The government has certain rights in the invention. TECHNICAL FIELD [0003] The present disclosure provides compositions for the prevention and treatment of genital herpes, comprising nucleoside modified RNAs that encode herpes simplex virus (HSV) glycoproteins, including those involved in virus entry and immune evasion, and methods of use thereof. BACKGROUND [0004] Herpes simplex type 1 (HSV-1) infections affect an estimated 3.7 billion people between the ages of 0-49 worldwide. Many people acquire HSV-1 during childhood from family members, friends, or peers at school. Orolabial HSV-1 infection is usually self-limited and although emotionally disturbing to some, does not pose a serious health risk to those infected. However, HSV-1 infection at other sites, such as the cornea or brain, may cause blindness and life- threatening encephalitis, respectively. In many resource-rich countries, reduced orolabial HSV-1 acquisition during childhood has resulted in adolescents and adults having a higher risk of acquiring first-time HSV-1 as a genital infection. Although genital HSV- 1 infections have a reduced risk of recurrences, genital transmission of HSV-1 from mother to newborn during labor 1 11979815v1 P-627574-PC and delivery can be devastating. HSV-1 neonatal herpes is more common than HSV-2 neonatal herpes in resource rich-countries. [0005] A prophylactic vaccine that prevents genital HSV-1 and HSV-2 infection and HSV-1 at non-genital sites is a public health priority. Despite major efforts from the scientific community, no FDA-approved vaccine is available to prevent HSV-1 or HSV-2 infection. SUMMARY [0006] The present disclosure provides pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) for delivering a particular herpes simplex virus (HSV) glycoprotein or immunogenic fragment thereof (e.g., HSV-1 glycoprotein or immunogenic fragment thereof, an HSV-2 glycoprotein or immunogenic fragment thereof, or any combinations thereof) to a subject (e.g., a patient) and related technologies (e.g., methods). In particular, the present disclosure provides HSV (e.g., HSV-1, HSV-2, or both) vaccine compositions and related technologies (e.g., methods). Additionally, the present disclosure provides HSV (e.g., HSV-1, HSV-2, or both) vaccine compositions for inhibiting, preventing or treating an HSV infection (e.g., HSV-1, HSV-2, or both). [0007] In some embodiments, the present disclosure provides a method of inhibiting an HSV-1 oral mucosal infection in a subject comprising the step of administering to said subject a composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, a composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and a composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof. [0008] In another embodiment, the present disclosure provides a method of preventing establishment of a latent HSV infection in a subject comprising the step of administering to said subject a composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, a composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and a composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof. [0009] Other features and advantages of the present disclosure will become apparent from the following detailed description examples and figures. It should be understood, however, that the detailed description and the specific examples while indicating preferred embodiments of the disclosure are given by way of illustration only, since various changes and modifications within 2 11979815v1 P-627574-PC the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description. BRIEF DESCRIPTION OF THE DRAWINGS [0010] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present disclosure, the inventions of which can be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein. The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. [0011] Figures 1A-1C. Characterization of exemplary translational products of an immunogenic fragment of the ectodomain from HSV-2 gC (gC2)-, HSV-2 gD (gD2)- and HSV-2 gE (gE2)- modified RNA in Vero cells. [0012] Figure 1A. Western blot showing expression of an immunogenic fragment of gC2 from an exemplary modified RNA. [0013] Figure 1B. Western blot showing expression of an immunogenic fragment of gD2 from an exemplary modified RNA. [0014] Figure 1C. Western blot showing expression of an immunogenic fragment of gE2 from an exemplary modified RNA. [0015] Figure 2A. gC2 antibody (Ab) response as determined by antigen-specific ELISA in mice immunized with an exemplary gD2 RNA; or an exemplary combination of a gC2 RNA, a gD2 RNA and a gE2 RNA each given at a different intradermal site (Trivalent-I); or an exemplary combination of a gC2 RNA, a gD2 RNA and a gE2 RNA given in a single composition (Trivalent- C). I indicates first immunization; II indicates second immunization. [0016] Figure 2B. gD2 Ab response as determined by antigen-specific ELISA in mice immunized with an exemplary gD2 RNA; or an exemplary combination of a gC2 RNA, a gD2 RNA and a gE2 RNA each given at a different intradermal site (Trivalent-I); or an exemplary combination of a gC2 RNA, a gD2 RNA and a gE2 RNA given in a single composition (Trivalent- C). I indicates first immunization; II indicates second immunization. [0017] Figure 2C. gE2 Ab response as determined by antigen-specific ELISA in mice immunized with an exemplary gD2 RNA; or an exemplary combination of a gC2 RNA, a gD2 RNA and a gE2 RNA each given at a different intradermal site (Trivalent-I); or an exemplary 3 11979815v1 P-627574-PC combination of a gC2 RNA, gD2 RNA and gE2 RNA given in a single composition (Trivalent- C). I indicates first immunization; II indicates second immunization. [0018] Figure 3A. Antigen-specific IgG1 responses in mice vaccinated with exemplary RNA compositions. Antibodies were evaluated after the first and second immunization for IgG1 responses. I indicates first immunization; II indicates second immunization. [0019] Figure 3B. Antigen-specific IgG2a responses in mice vaccinated with exemplary RNA compositions. Antibodies were evaluated after the first and second immunization for IgG2a responses. I indicates first immunization; II indicates second immunization. [0020] Figure 4. Neutralizing antibody titers in RNA vaccinated mice. 50% endpoint neutralization titers of serum were obtained after the second immunization. Titers were performed using 10% human complement. Trivalent-I animals were immunized with an exemplary gC2/lipid nanoparticle (LNP), an exemplary gD2/LNP, and an exemplary gE2/LNP, each given at a different site. Trivalent-C animals were immunized with an exemplary gC2, an exemplary gD2 and an exemplary gE2 combined into a single LNP. P values comparing 50% endpoint neutralizing titers: Trivalent-I versus gD2, p=0.04; Trivalent-C versus gD2, p=0.002; Trivalent-I versus Trivalent-C, p-0,026. [0021] Figures 5A-5B. CD4+ T cell responses to an exemplary gC2, an exemplary gD2 and an exemplary gE2 RNA each administered at a different intradermal site. Splenocytes were stimulated with subunit antigen glycoproteins (Figure 5A) or 15 amino acid peptides with 11 overlapping amino acids to stimulate HSV-2 specific T cell responses (Figure 5B). * indicates p<0.05 (t test) comparing gC, gD or gE with PBS stimulated CD4+ T cells or DMSO stimulated CD4+ T cells. Error bars represent SD. [0022] Figures 6A-6B. CD8+ T cell responses to an exemplary gC2, an exemplary gD2 and an exemplary gE2 RNA, each administered at a different intradermal site. Splenocytes were stimulated with subunit antigen glycoproteins (Figure 6A) or 15 amino acid peptides with 11 overlapping amino acids to stimulate HSV-2 specific T cell responses (Figure B). * indicates p<0.05 comparing gE pool 2 with DMSO control. Error bars represent SD. [0023] Figure 7A. Survival in BALB/c mice immunized with exemplary RNA compositions twice at 28 day intervals and challenged intravaginally with HSV-2. Trivalent-I represents animals immunized with an exemplary gC2 RNA/LNP, an exemplary gD2 RNA /LNP, and an exemplary gE2 RNA /LNP each given at different intradermal sites. Trivalent-C represents 4 11979815v1 P-627574-PC animals immunized with an exemplary gC2 RNA, an exemplary gD2 RNA and an exemplary gE2 RNA combined into a single LNP for immunization. [0024] Figure 7B. Weight loss (-) or gain (+) and neurological signs in BALB/c mice immunized with exemplary RNA compositions twice at 28 day intervals and challenged intravaginally with HSV-2. Trivalent-I represents animals immunized with an exemplary gC2 RNA/LNP, an exemplary gD2 RNA /LNP, and an exemplary gE2 RNA /LNP each given at different intradermal sites. Trivalent-C represents animals immunized with an exemplary gC2 RNA, an exemplary gD2 RNA and an exemplary gE2 RNA combined into a single LNP for immunization. [0025] Figures 8A-8B. Vaginal viral titers in mice vaccinated with exemplary RNA compositions after intravaginal challenge with HSV-2. Vaginal swab titers were obtained on Day 2 (Figure 8A) and Day 4 (Figure 8B) post-challenge. The dotted lines indicate the limit of detection of the assay at 7 PFU/ml. [0026] Figure 9. Genital disease in mice vaccinated with exemplary RNA compositions after HSV-2 vaginal challenge. Mice were immunized with poly C as a control, or with an exemplary gD2 RNA/LNP, trivalent at individual sites for each exemplary glycoprotein RNA (Trivalent-I) or trivalent with all three exemplary RNAs combined (Trivalent-C). Genital disease was scored for 28 days. All animals in the poly C group died by day 10. ***, indicates p<0.001 comparing poly C with the other 3 groups. [0027] Figure 10. HSV-2 DNA copies in dorsal root ganglia (DRG) in RNA vaccinated mice on day 4 post challenge. HSV-2 DNA in DRG was measured by qPCR. DRG from 4 to 5 animals per group were evaluated for HSV-2 DNA at 4 days post challenge. The bars represent the mean values per group. [0028] Figure 11A. The exemplary trivalent RNA-LNP vaccine induces a potent T follicular helper cell response in mice. BALB/c female mice were left un-immunized as naïve control animals or immunized intradermally twice at 28 day intervals with poly C RNA-LNP or an exemplary trivalent modified RNA-LNP. The poly C RNA controls received 10μg Poly C RNA- LNP divided into 4 aliquots and administered at 4 separate sites. The exemplary trivalent modified RNA group received 10μg of an exemplary gC2 RNA-LNP, 10μg of an exemplary gD2 RNA- LNP, and 10μg of an exemplary gE2 RNA-LNP each divided into 2 aliquots and each given at 2 sites. Two weeks after the second immunization, spleens were harvested from 5 animals per group and flow cytometry performed to detect T follicular helper (Tfh) cell responses (*p<0.05). 5 11979815v1 P-627574-PC [0029] Figure 11B. The exemplary trivalent RNA-LNP vaccine induces a potent germinal center B cell response in mice. BALB/c female mice were left un-immunized as naïve control animals or immunized intradermally twice at 28 day intervals with poly C RNA-LNP or an exemplary trivalent modified RNA-LNP. The poly C RNA controls received 10μg Poly C RNA- LNP divided into 4 aliquots and administered at 4 separate sites. The trivalent modified RNA group received 10μg of an exemplary gC2 RNA-LNP, 10μg of an exemplary gD2 RNA-LNP, and 10μg of an exemplary gE2 RNA-LNP each divided into 2 aliquots and each given at 2 sites. Two weeks after the second immunization, spleens were harvested from 5 animals per group and flow cytometry performed to detect germinal center B cell responses (*p<0.05). [0030] Figures 12A-12C. Genital mucosa IgG antibody responses. BALB/c mice were immunized intradermally twice at 28 day intervals with 10μg of poly C RNA-LNP, 10μg of an exemplary gD2 RNA-LNP or 10μg each of an exemplary gC2, an exemplary gD2, an exemplary gE2 trivalent modified RNA-LNP. The exemplary trivalent RNA was combined and administered as 10μg of an exemplary gC2 RNA & 10μg of an exemplary gD2 RNA & 10μg of an exemplary gE2 RNA combined into LNP and divided into 4 aliquots and given at 4 sites. One month after the second immunization, 60μl of media was introduced in the vaginal cavity and retrieved. IgG titers were determined at a 1:50 dilution of the vaginal wash fluids by ELISA to gC2 (Figure 12A), gD2 (Figure 12B), and gE2 (Figure 12C) (n=10 mice in the poly C group, n=10 in the gD2 RNA group and n=25 in the trivalent RNA group; ***p<0.001; **p<0.01). [0031] Figure 13. The exemplary trivalent RNA-LNP vaccine produces antibodies that block gC2 binding to complement component C3b. BALB/c mice were left unimmunized as a source of non-immune IgG, or immunized intradermally with poly C RNA-LNP or an exemplary trivalent RNA-LNP. The poly C RNA controls received 10μg poly C RNA-LNP divided into 4 aliquots and administered at 4 separate sites. The gD2 RNA group received 10μg of an exemplary gD2 RNA- LNP administered as described for the poly C RNA-LNP. The trivalent modified RNA group received 10μg of an exemplary gC2 RNA-LNP, 10μg of an exemplary gD2 RNA-LNP, and 10μg of an exemplary gE2 RNA-LNP combined into LNP and divided into 4 aliquots and given at 4 sites. There were 10 mice in each group. Sera from the 10 mice were pooled and IgG was purified. The IgG was evaluated at 12μg/200μl for its ability to block complement component C3b binding to gC2. (****p<0.0001). [0032] Figures 14A-14F. The exemplary trivalent RNA vaccine provides outstanding protection in mice when the vaccine is administered intramuscularly. BALB/c mice were 6 11979815v1 P-627574-PC immunized intramuscularly with poly C RNA-LNP as a control (15/group) or with an exemplary trivalent RNA composition containing 10 μg each of an exemplary gC2, an exemplary gD2 and an exemplary gE2 RNA-LNP (20/group). Figure 14A presents data on mouse survival; Figure 14B presents data on weight loss; Figure 14C presents data on genital disease. DRG were harvested from nine poly C animals at the time of euthanasia between days 7 and 12 post-infection or at the end of the experiment on day 28 in the trivalent RNA group. Figure 14D presents data on HSV-2 DNA in DRG. Figure 14E presents data on vaginal viral cultures on Day 2 and Figure 14F presents data on vaginal viral cultures on Day 4. Difference between poly C and trivalent groups are significant, p<0.001 for Figures 14A-14F. [0033] Figures 15A-15C. The exemplary trivalent RNA vaccine is highly efficacious in the guinea pig genital infection model. Hartley Strain female guinea pigs were left unimmunized and uninfected (naive group, n=10), immunized three times intradermally at monthly intervals with 20μg poly C RNA-LNP (n=10) or with 20μg each of an exemplary gC2, an exemplary gD2, an exemplary gE modified RNA-LNP (n=10). One month after the final immunization, animals in the poly C and trivalent RNA groups were infected intravaginally with 5x105 PFU of HSV-2 strain MS (50 LD50). Animals were observed for death, genital lesions during the acute phase of infection (days 1-14) and genital lesions during the recurrent phase of infection (days 15-60). Figure 15A presents data on survival; Figure 15B provides data on vaginal disease (acute phase); and Figure 15C provides data on vaginal disease (recurrent phase). [0034] Figures 16A-16D. Exemplary tri-HSV-1 and exemplary tri-HSV-2 immunizations generate antibodies that bind to HSV-1 proteins and neutralize HSV-1. Mice were immunized twice with PBS, exemplary tri-HSV-1 RNA, or exemplary tri-HSV-2 RNA at 1 µg or 10 µg of total RNA. Figures 16A-16C. IgG ELISA endpoint titers against HSV-1 proteins. Figure 16D. HSV-1 neutralizing titers in the presence of 5% human serum as source of complement. N=10 mice per group. P values were calculated using the two-tailed Mann-Whitney test. [0035] Figure 17. Gating strategy used in CD4 and CD8 T cell flow cytometry analysis. Singlet cells were gated, then live CD3+, then lymphocytes, and then CD4+ and CD8+ T-cells. Single cytokine positive CD8+ T-cells were gated using CD8+ and TNFα, IFNγ, or IL-2 parameters. Gate coordinates were set based on the DMSO treated cells and those gate coordinates were transferred to the peptide treated cells. [0036] Figures 18A-18D. Exemplary tri-HSV-1 and exemplary tri-HSV-2 immunizations generate CD4+ and CD8+ T-cell responses to HSV-1 overlapping peptides. Mice were 7 11979815v1 P-627574-PC immunized twice with 10 µg total of exemplary tri-HSV-1 RNA, exemplary tri-HSV-2 RNA, or PBS. Figure 18A. Proportion of CD8+ T-cells producing 0, 1, 2, or 3 cytokines. Numbers inside or adjacent to colored segments indicate percentage of total CD8+ T-cells producing the indicated number of cytokines. Figure 18B. Activation markers of CD8+ T-cells stimulated with gE1 overlapping peptide pool producing TNFα, IFNγ, and/or IL-2. Figure 18C. Proportion of CD8+ T-cells expressing CD107a when stimulated with gE1 overlapping peptides. Figure 18D. Proportion of CD4+ T-cells producing polyfunctional cytokines TNFα, IFNγ, and IL-2 following stimulation with a gD1 overlapping peptide pool. N=5 mice per group. P values in Figure 18B were calculated using 2-way ANOVA with Tukey’s correction for multiple comparisons. P values in Figure 18C and Figure 18D were calculated using the two-tailed Mann-Whitney test. [0037] Figures 19A-19G. Exemplary tri-HSV-1 and tri-HSV-2 RNA compositions protect mice against acute and latent HSV-1 infection in the lip. Mice were immunized with PBS, exemplary tri-HSV-1 RNA, or tri-HSV-2 RNA compositions as in Figure 1. Mice were challenged with 2 ×106 PFU of HSV-1 applied to the scarified lower lip. Figure 19A. HSV-1 virus titers in the lower lip. Figure 19B. HSV-1 DNA copy number in the lower lip. Figure 19C. HSV-1 virus titers in the trigeminal ganglia (TG). Figure 19D. HSV-1 DNA copy number in the TG. Figures 19A-19D. n=10 per group. Figure 19E. Mice were weighed daily for 13 days. Figure 19F. Mice were scored for lesions on the lip with a score of 1 for lesions or 0 for no lesions. Figure 19G. HSV-1 DNA genome copies in the TG 28 days post-challenge. Figures 19E-19G. n=24 mice for PBS group, n=15 mice per RNA immunization group. P values in Figures 19A-19D, and Figure 19G were calculated by the two-tailed Mann-Whitney test. [0038] Figures 20A-20F. Exemplary tri-HSV-1 and exemplary tri-HSV-2 RNA compositions protect mice from intravaginal HSV-1 disease. Mice were immunized with PBS, exemplary tri- HSV-1 RNA, or exemplary tri-HSV-2 RNA. Mice were challenged with 2 ×106 PFU of HSV-1 intravaginally and scored for clinical disease for 14 days. Figure 20A. Survival curve. P values were calculated by the log-rank test. Figure 20B. Weight loss. Figure 20C. Mean genital disease scores for each group. Figures 20D-20E. Vaginal swabs were obtained on days two and four for virus titers. Figure 20F. HSV-1 DNA copy number in DRG. N=5 mice for PBS group, n=5 mice for tri-HSV-1 group, and n=4 mice for tri-HSV-2 group. P values in Figures 20D-20F were calculated by the two-tailed Mann-Whitney test without adjustments for multiple comparisons. [0039] Figure 21. Determination of LD50 of HSV-1. Naïve (not vaccinated) 6 to 8-week-old female mice were inoculated intranasally with 5x103, 5x104, 5x105, 5x106 plaque-forming units 8 11979815v1 P-627574-PC (pfu) of HSV-1 strain H129, or with PBS as a control. Survival was evaluated over 28 days. Survival was 50% (2 of 4 mice) at 5x103 pfu, establishing the 50% lethal dose (LD50) as 5x103 pfu. [0040] Figures 22A-22D. Trivalent HSV-2 vaccine (Tri-HSV-2) protects BALB/c mice against HSV-1 encephalitis. Female BALB/c mice were immunized twice with either 1 ^g (0.33 ^g of each mRNA) or 10 ^g (3.33 ^g each mRNA) trivalent HSV-2 mRNA vaccine at 1-month intervals or PBS as described above. One month after the 2nd immunization, animals were infected intranasally with 5x105 pfu (100 LD50) of HSV-1 strain H129. Figure 22A. Animals were monitored for survival; P<0.01 comparing 1 or 10 ^g Tri-HSV-2 vaccine group with PBS by Log- rank test. Figures 22B-22D. HSV-1 DNA copy number was determined on day 6 post-infection by qPCR in brains (Figure 22B), trigeminal ganglia (Figure 22C), and olfactory bulbs (Figure 22D); n=10/group except n=9 for olfactory bulb in the 1 ^g Tri-HSV-2 vaccine group. P value comparing log10 copy number (within the graph): ***, P<0.001; P values were calculated by two- tailed Mann-Whitney test. P value comparing number of animals positive for HSV-1 DNA (above the graph): **, P<0.01; P values were calculated by two-tailed Fisher’s exact test. CERTAIN DEFINITIONS [0041] In general, terminology used herein is in accordance with its understood meaning in the art, unless clearly indicated otherwise. Explicit definitions of certain terms are provided below; meanings of these and other terms in particular instances throughout this specification will be clear to those skilled in the art from context. [0042] In order that the present invention may be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification. [0043] About: The term “about”, when used herein in reference to a value, refers to a value that is similar, in context to the referenced value. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variance encompassed by “about” in that context. For example, in some embodiments, the term “about” may encompass a range of values that within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value. [0044] Agent: As used herein, the term “agent”, may refer to a physical entity or phenomenon. In some embodiments, an agent may be characterized by a particular feature and/or effect. In some 9 11979815v1 P-627574-PC embodiments, an agent may be a compound, molecule, or entity of any chemical class including, for example, a small molecule, polypeptide, nucleic acid, saccharide, lipid, metal, or a combination or complex thereof. In some embodiments, the term “agent” may refer to a compound, molecule, or entity that comprises a polymer. In some embodiments, the term may refer to a compound or entity that comprises one or more polymeric moieties. In some embodiments, the term “agent” may refer to a compound, molecule, or entity that is substantially free of a particular polymer or polymeric moiety. In some embodiments, the term may refer to a compound, molecule, or entity that lacks or is substantially free of any polymer or polymeric moiety. [0045] Amino acid: In its broadest sense, as used herein, the term “amino acid” refers to a compound and/or substance that can be, is, or has been incorporated into a polypeptide chain, e.g., through formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N–C(H)(R)–COOH. In some embodiments, an amino acid is a naturally- occurring amino acid. In some embodiments, an amino acid is a non-natural amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L- amino acid. “Standard amino acid” refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. “Nonstandard amino acid” refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. In some embodiments, an amino acid, including a carboxy- and/or amino-terminal amino acid in a polypeptide, can contain a structural modification as compared with the general structure above. For example, in some embodiments, an amino acid may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and/or substitution (e.g., of the amino group, the carboxylic acid group, one or more protons, and/or the hydroxyl group) as compared with the general structure. In some embodiments, such modification may, for example, alter the circulating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino acid. In some embodiments, such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino acid, as compared with one containing an otherwise identical unmodified amino acid. As will be clear from context, in some embodiments, the term “amino acid” may be used to refer to a free amino acid; in some embodiments it may be used to refer to an amino acid residue of a polypeptide. [0046] Antibody agent: As used herein, the term “antibody agent” refers to an agent that specifically binds to a particular antigen. In some embodiments, the term encompasses a 10 11979815v1 P-627574-PC polypeptide or polypeptide complex that includes immunoglobulin structural elements sufficient to confer specific binding. For example, in some embodiments, an antibody agent is or comprises a polypeptide whose amino acid sequence includes one or more structural elements recognized by those skilled in the art as a complementarity determining region (CDR); in some embodiments an antibody agent is or comprises a polypeptide whose amino acid sequence includes at least one CDR (e.g., at least one heavy chain CDR and/or at least one light chain CDR) that is substantially identical to one found in a reference antibody. In some embodiments an included CDR is substantially identical to a reference CDR in that it is either identical in sequence or contains between 1-5 amino acid substitutions as compared with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that it shows at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that it shows at least 96%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that at least one amino acid within the included CDR is deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that 1-5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that at least one amino acid within the included CDR is substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that 1-5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments, an antibody agent is or comprises a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art as an immunoglobulin variable domain. In some embodiments, an antibody agent in or comprises a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art to correspond to CDRs1, 2, and 3 of an antibody variable domain; in some such embodiments, an antibody agent in or comprises a polypeptide or set of polypeptides whose amino acid sequence(s) 11 11979815v1 P-627574-PC together include structural elements recognized by those skilled in the art to correspond to both heavy chain and light chain variable region CDRs, e.g., heavy chain CDRs 1, 2, and/or 3 and light chain CDRs 1, 2, and/or 3. In some embodiments, an antibody agent is a polypeptide protein having a binding domain which is homologous or largely homologous to an immunoglobulin- binding domain. In some embodiments, an antibody agent may be or comprise a polyclonal antibody preparation. In some embodiments, an antibody agent may be or comprise a monoclonal antibody preparation. In some embodiments, an antibody agent may include one or more constant region sequences that are characteristic of a particular organism, such as a camel, human, mouse, primate, rabbit, rat; in many embodiments, an antibody agent may include one or more constant region sequences that are characteristic of a human. In some embodiments, an antibody agent may include one or more sequence elements that would be recognized by one skilled in the art as a humanized sequence, a primatized sequence, a chimeric sequence, etc. In some embodiments, an antibody agent may be a canonical antibody (e.g., may comprise two heavy chains and two light chains). In some embodiments, an antibody agent may be in a format selected from, but not limited to, intact IgA, IgG, IgE or IgM antibodies; bi- or multi- specific antibodies (e.g., Zybodies®, etc); antibody fragments such as Fab fragments, Fab’ fragments, F(ab’)2 fragments, Fd’ fragments, Fd fragments, and isolated CDRs or sets thereof; single chain Fvs; polypeptide-Fc fusions; single domain antibodies (e.g., shark single domain antibodies such as IgNAR or fragments thereof); cameloid antibodies; masked antibodies (e.g., Probodies®); Small Modular ImmunoPharmaceuticals (“SMIPsTM”); single chain or Tandem diabodies (TandAb®); VHHs; Anticalins®; Nanobodies® minibodies; BiTE®s; ankyrin repeat proteins or DARPINs®; Avimers®; DARTs; TCR-like antibodies;, Adnectins®; Affilins®; Trans-bodies®; Affibodies®; TrimerX®; MicroProteins; Fynomers®, Centyrins®; and KALBITOR®s. In some embodiments, an antibody may lack a covalent modification (e.g., attachment of a glycan) that it would have if produced naturally. In some embodiments, an antibody may contain a covalent modification (e.g., attachment of a glycan, a payload (e.g., a detectable moiety, a therapeutic moiety, a catalytic moiety, etc.), or other pendant group (e.g., poly-ethylene glycol, etc.)). [0047] Antigen: Those skilled in the art, reading the present specification, will appreciate that the term “antigen” refers to a molecule that is recognized by the immune system, e.g., in particular embodiments the adaptive immune system, such that it elicits an antigen-specific immune response. In some embodiments, an antigen-specific immune response may be or comprise generation of antibodies and/or antigen-specific T cells. In some embodiments, an antigen is a 12 11979815v1 P-627574-PC peptide or polypeptide that comprises at least one epitope against which an immune response can be generated. In some embodiments, an antigen is presented by cells of the immune system such as antigen presenting cells like dendritic cells or macrophages. In one embodiments, an antigen or a processed product thereof such as a T-cell epitope is bound by a T- or B-cell receptor, or by an immunoglobulin molecule such as an antibody. Accordingly, an antigen or a processed product thereof may react specifically with antibodies or T lymphocytes (T cells). In some embodiments, an antigen is a parasitic antigen. In accordance with the present disclosure, in some embodiments, an antigen may be delivered by RNA molecules as described herein. In some embodiments, a peptide or polypeptide antigen can be 2-100 amino acids, including for example, 5 amino acids, 10 amino acids, 15 amino acids, 20 amino acids, 25 amino acids, 30 amino acids, 35 amino acids, 40 amino acids, 45 amino acids, or 50 amino acids in length. In some embodiments, a peptide or polypeptide antigen can be greater than 50 amino acids. In some embodiments, a peptide or polypeptide antigen can be greater than 100 amino acids. In some embodiments, an antigen is recognized by an immune effector cell. In some embodiments, an antigen if recognized by an immune effector cell is able to induce in the presence of appropriate co-stimulatory signals, stimulation, priming and/or expansion of the immune effector cell carrying an antigen receptor recognizing the antigen. In the context of the embodiments of the present disclosure, in some embodiments, an antigen can be presented or present on the surface of a cell, e.g., an antigen presenting cell. In some embodiments, an antigen is presented by a diseased cell such as a virus- infected cell. In some embodiments, an antigen receptor is a TCR which binds to an epitope of an antigen presented in the context of MHC. In some embodiments, binding of a TCR when expressed by T cells and/or present on T cells to an antigen presented by cells such as antigen presenting cells results in stimulation, priming and/or expansion of said T cells. In some embodiments, binding of a TCR when expressed by T cells and/or present on T cells to an antigen presented on diseased cells results in cytolysis and/or apoptosis of the diseased cells, wherein said T cells preferably release cytotoxic factors, e.g., perforins and granzymes. [0048] Associated: Two events or entities are “associated” with one another, as that term is used herein, if the presence, level, degree, type and/or form of one is correlated with that of the other. For example, a particular entity (e.g., polypeptide, genetic signature, metabolite, microbe, etc) is considered to be associated with a particular disease, disorder, or condition, if its presence, level and/or form correlates with incidence of, susceptibility to, severity of, stage of, etc. the disease, disorder, or condition (e.g., across a relevant population). In some embodiments, two or more 13 11979815v1 P-627574-PC entities are physically “associated” with one another if they interact, directly or indirectly, so that they are and/or remain in physical proximity with one another. In some embodiments, two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non-covalently associated, for example by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof. [0049] Binding: Those skilled in the art, reading the present specification, will appreciate that the term “binding” typically refers to a non-covalent association between or among entities or moieties. In some embodiments, binding data are expressed in terms of “IC50”. As is understood in the art, IC50 is the concentration of an assessed agent in a binding assay at which 50% inhibition of binding of reference agent known to bind the relevant binding partner is observed. In some embodiments, assays are run under conditions in which the assays are run (e.g., limiting binding target and reference concentrations), these values approximate KD values. Assays for determining binding are well known in the art and are described in detail, for example, in PCT publications WO 94/20127 and WO 94/03205, and other publications such Sidney et al., Current Protocols in Immunology 18.3.1 (1998); Sidney, et al., J. Immunol. 154:247 (1995); and Sette, et al., Mol. Immunol.31:813 (1994). Alternatively, binding can be expressed relative to binding by a reference standard peptide. For example, can be based on its IC50, relative to the IC50 of a reference standard peptide. Binding can also be determined using other assay systems including those using: live cells (e.g., Ceppellini et al., Nature 339:392 (1989); Christnick et al., Nature 352:67 (1991); Busch et al., Int. Immunol. 2:443 (1990); Hill et al., J. Immunol. 147:189 (1991); del Guercio et al., J. Immunol. 154:685 (1995)), cell free systems using detergent lysates (e.g., Cerundolo et al., J. Immunol 21:2069 (1991)), immobilized purified MHC (e.g., Hill et al., J. Immunol. 152, 2890 (1994); Marshall et al., J. Immunol. 152:4946 (1994)), ELISA systems (e.g., Reay et al., EMBO J. 11:2829 (1992)), surface plasmon resonance (e.g., Khilko et al., J. Biol. Chem. 268:15425 (1993)); high flux soluble phase assays (Hammer et al., J. Exp. Med. 180:2353 (1994)), and measurement of class I MHC stabilization or assembly (e.g., Ljunggren et al., Nature 346:476 (1990); Schumacher et al., Cell 62:563 (1990); Townsend et al., Cell 62:285 (1990); Parker et al., J. Immunol.149:1896 (1992)). [0050] Cap: As used herein, the term “cap” refers to a structure comprising or essentially consisting of a nucleoside-5 '-triphosphate that is typically joined to a 5'-end of an uncapped RNA 14 11979815v1 P-627574-PC (e.g., an uncapped RNA having a 5'- diphosphate). In some embodiments, a cap is or comprises a guanine nucleotide. In some embodiments, a cap is or comprises a naturally-occurring RNA 5’ cap, including, e.g., but not limited to a 7- methylguanosine cap, which has a structure designated as “m7G.” In some embodiments, a cap is or comprises a synthetic cap analog that resembles an RNA cap structure and possesses the ability to stabilize RNA if attached thereto, including, e.g., but not limited to anti-reverse cap analogs (ARCAs) known in the art). Those skilled in the art will appreciate that methods for joining a cap to a 5’ end of an RNA are known in the art. For example, in some embodiments, a capped RNA may be obtained by in vitro capping of RNA that has a 5' triphosphate group or RNA that has a 5' diphosphate group with a capping enzyme system (including, e.g., but not limited to vaccinia capping enzyme system or Saccharomyces cerevisiae capping enzyme system). Alternatively, a capped RNA can be obtained by in vitro transcription (IVT) of a single-stranded DNA template in the presence of a dinucleotide or trinucleotide cap analog. [0051] Cell-mediated immunity: “Cell-mediated immunity,” “cellular immunity,” “cellular immune response,” or similar terms are meant to include a cellular response directed to cells characterized by expression of an antigen, in particular characterized by presentation of an antigen with class I or class II MHC. A cellular response relates to immune effector cells, in particular to T cells or T lymphocytes which act as either “helpers” or “killers.” The helper T cells (also termed CD4+ T cells or CD4 T cells) play a central role by regulating the immune response and the killer cells (also termed cytotoxic T cells, cytolytic T cells, CD8+ T cells, CD8 T cells, or CTLs) kill diseased cells such as virus-infected cells, preventing the production of more diseased cells. [0052] Co-administration: As used herein, the term “co-administration” refers to use of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) described herein and an additional therapeutic agent. The combined use of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) described herein and an additional therapeutic agent may be performed concurrently or separately (e.g., sequentially in any order). In some embodiments, a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) described herein and an additional therapeutic agent may be combined in one pharmaceutically-acceptable carrier, or they may be placed in separate carriers and delivered to a target cell or administered to a subject at different times. Each of these situations is contemplated as falling within the meaning of “co- administration” or “combination,” provided that a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) described herein and an additional therapeutic agent are delivered or 15 11979815v1 P-627574-PC administered sufficiently close in time that there is at least some temporal overlap in biological effect(s) generated by each on a target cell or a subject being treated. [0053] Codon-optimized: As used herein, the term “codon-optimized” refers to alteration of codons in a coding region of a nucleic acid molecule to reflect the typical codon usage of a host organism without preferably altering the amino acid sequence encoded by the nucleic acid molecule. Within the context of the present disclosure, in some embodiments coding regions are codon-optimized for optimal expression in a subject to be treated using the RNA molecules described herein. In some embodiments, codon-optimization may be performed such that codons for which frequently occurring tRNAs are available are inserted in place of “rare codons.” In some embodiments, codon-optimization may include increasing guanosine/cytosine (G/C) content of a coding region of RNA described herein as compared to the G/C content of the corresponding coding sequence of a wild type RNA, wherein the amino acid sequence encoded by the RNA is preferably not modified compared to the amino acid sequence. [0054] Combination therapy: As used herein, the term “combination therapy” refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents). In some embodiments, the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all “doses” of a first regimen are administered prior to administration of any doses of a second regimen); in some embodiments, such agents are administered in overlapping dosing regimens. In some embodiments, “administration” of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modality(ies) in the combination. For clarity, combination therapy does not require that individual agents be administered together in a single composition (or even necessarily at the same time), although in some embodiments, two or more agents, or active moieties thereof, may be administered together in a combination composition. [0055] Comparable: As used herein, the term “comparable” refers to two or more agents, entities, situations, sets of conditions, etc., that may not be identical to one another but that are sufficiently similar to permit comparison there between so that one skilled in the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed. In some embodiments, comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features. Those of ordinary skill in the art will understand, in context, what degree of identity is 16 11979815v1 P-627574-PC required in any given circumstance for two or more such agents, entities, situations, sets of conditions, etc. to be considered comparable. For example, those of ordinary skill in the art will appreciate that sets of circumstances, individuals, or populations are comparable to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by or indicative of the variation in those features that are varied. [0056] Corresponding to: As used herein, the term “corresponding to” refers to a relationship between two or more entities. For example, the term “corresponding to” may be used to designate the position/identity of a structural element in a compound or composition relative to another compound or composition (e.g., to an appropriate reference compound or composition). For example, in some embodiments, a monomeric residue in a polymer (e.g., an amino acid residue in a polypeptide or a nucleic acid residue in a polynucleotide) may be identified as “corresponding to” a residue in an appropriate reference polymer. For example, those of ordinary skill will appreciate that, for purposes of simplicity, residues in a polypeptide are often designated using a canonical numbering system based on a reference related polypeptide, so that an amino acid “corresponding to” a residue at position 190, for example, need not actually be the 190th amino acid in a particular amino acid chain but rather corresponds to the residue found at 190 in the reference polypeptide; those of ordinary skill in the art readily appreciate how to identify “corresponding” amino acids. For example, those skilled in the art will be aware of various sequence alignment strategies, including software programs such as, for example, BLAST, CS- BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH/GLSEARCH, Genoogle, HMMER, HHpred/HHsearch, IDF, Infernal, KLAST, USEARCH, parasail, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE that can be utilized, for example, to identify “corresponding” residues in polypeptides and/or nucleic acids in accordance with the present disclosure. Those of skill in the art will also appreciate that, in some instances, the term “corresponding to” may be used to describe an event or entity that shares a relevant similarity with another event or entity (e.g., an appropriate reference event or entity). To give but one example, a gene or protein in one organism may be described as “corresponding to” a gene or protein from another organism in order to indicate, in some embodiments, that it plays an analogous role or performs an analogous function and/or that it shows a particular degree of sequence identity or homology, or shares a particular characteristic sequence element. 17 11979815v1 P-627574-PC [0057] Derived: In the context of an amino acid sequence (peptide or polypeptide) “derived from” a designated amino acid sequence (peptide or polypeptide), it refers to a structural analogue of a designated amino acid sequence. In some embodiments, an amino acid sequence which is derived from a particular amino acid sequence has an amino acid sequence that is identical, essentially identical or homologous to that particular sequence or a fragment thereof. Amino acid sequences derived from a particular amino acid sequence may be variants of that particular sequence or a fragment thereof. For example, it will be understood by one of ordinary skill in the art that the antigens suitable for use herein may be altered such that they vary in sequence from the naturally occurring or native sequences from which they were derived, while retaining the desirable activity of the native sequences. [0058] Designed: As used herein, the term “designed” refers to an agent (i) whose structure is or was selected by the hand of man; (ii) that is produced by a process requiring the hand of man; and/or (iii) that is distinct from natural substances and other known agents. [0059] Dosing regimen: Those skilled in the art will appreciate that the term “dosing regimen” may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses. In some embodiments, a dosing regimen comprises a plurality of doses each of which is separated in time from other doses. In some embodiments, individual doses are separated from one another by a time period of the same length; in some embodiments, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen). [0060] Engineered: In general, the term “engineered” refers to the aspect of having been manipulated by the hand of man. For example, a polynucleotide is considered to be “engineered” when two or more sequences that are not linked together in that order in nature are manipulated 18 11979815v1 P-627574-PC by the hand of man to be directly linked to one another in the engineered polynucleotide and/or when a particular residue in a polynucleotide is non-naturally occurring and/or is caused through action of the hand of man to be linked with an entity or moiety with which it is not linked in nature. [0061] Epitope: As used herein, the term “epitope” refers to a moiety that is specifically recognized by an immunoglobulin (e.g., antibody or receptor) binding component. For example, an epitope may be recognized by a T cell, a B cell, or an antibody. In some embodiments, an epitope is comprised of a plurality of chemical atoms or groups on an antigen. In some embodiments, such chemical atoms or groups are surface-exposed when the antigen adopts a relevant three-dimensional conformation. In some embodiments, such chemical atoms or groups are physically near to each other in space when the antigen adopts such a conformation. In some embodiments, at least some such chemical atoms are groups are physically separated from one another when the antigen adopts an alternative conformation (e.g., is linearized). Accordingly, in some embodiments, an epitope of an antigen may include a continuous or discontinuous portion of the antigen. In some embodiments, an epitope is or comprises a T cell epitope. In some embodiments, an epitope may have a length of about 5 to about 30 amino acids, or about 10 to about 25 amino acids, or about 5 to about 15 amino acids, or about 5 to 12 amino acids, or about 6 to about 9 amino acids. [0062] Expression: As used herein, the term “expression” of a nucleic acid sequence refers to the generation of a gene product from the nucleic acid sequence. In some embodiments, a gene product can be a transcript. In some embodiments, a gene product can be a polypeptide. In some embodiments, expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, etc); (3) translation of an RNA into a polypeptide or protein; and/or (4) post-translational modification of a polypeptide or protein. [0063] Five prime untranslated region: As used herein, the terms “five prime untranslated region” or “5' UTR” refer to a sequence of an RNA molecule between a transcription start site and a start codon of a coding region of an RNA. In some embodiments, “5’ UTR” refers to a sequence of an RNA molecule that begins at a transcription start site and ends one nucleotide (nt) before a start codon (usually AUG) of a coding region of an RNA molecule, e.g., in its natural context. [0064] Humoral immunity: As used herein, the term “humoral immunity” or “humoral immune response” refers to antibody production and the accessory processes that accompany it, including: Th2 activation and cytokine production, germinal center formation and isotype switching, affinity 19 11979815v1 P-627574-PC maturation and memory cell generation. It also refers to the effector functions of antibodies, which include pathogen neutralization, classical complement activation, and opsonin promotion of phagocytosis and pathogen elimination. [0065] Identity: As used herein, the term “identity” refers to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules are considered to be “substantially identical” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical. Calculation of the percent identity of two nucleic acid or polypeptide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequence for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of a reference sequence. The nucleotides at corresponding positions are then compared. When a position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which has been incorporated into the ALIGN program (version 2.0). In some exemplary embodiments, nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix. [0066] Immunologically equivalent: The term “immunologically equivalent” means that an immunologically equivalent molecule such as the immunologically equivalent amino acid sequence exhibits the same or essentially the same immunological properties and/or exerts the 20 11979815v1 P-627574-PC same or essentially the same immunological effects, e.g., with respect to the type of the immunological effect. In the context of the present disclosure, in some embodiments, the term “immunologically equivalent” is used with respect to the immunological effects or properties of antigens or antigen variants used for immunization. For example, an amino acid sequence is immunologically equivalent to a reference amino acid sequence if said amino acid sequence when exposed to the immune system of a subject induces an immune reaction having a specificity of reacting with the reference amino acid sequence. [0067] In some embodiments, an antigen receptor is an antibody or B cell receptor which binds to an epitope in an antigen. In some embodiments, an antibody or B cell receptor binds to native epitopes of an antigen. [0068] Increased, Induced, or Reduced: As used herein, these terms or grammatically comparable comparative terms, indicate values that are relative to a comparable reference measurement. For example, in some embodiments, an assessed value achieved with a provided pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) may be “increased” relative to that obtained with a comparable reference pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine). Alternatively or additionally, in some embodiments, an assessed value achieved in a subject may be “increased” relative to that obtained in the same subject under different conditions (e.g., prior to or after an event; or presence or absence of an event such as administration of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) as described herein, or in a different, comparable subject (e.g., in a comparable subject that differs from the subject of interest in prior exposure to a condition, e.g., absence of administration of a pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) as described herein.). In some embodiments, comparative terms refer to statistically relevant differences (e.g., that are of a prevalence and/or magnitude sufficient to achieve statistical relevance). Those skilled in the art will be aware, or will readily be able to determine, in a given context, a degree and/or prevalence of difference that is required or sufficient to achieve such statistical significance. In some embodiments, the term “reduced” or equivalent terms refers to a reduction in the level of an assessed value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75% or higher, as compared to a comparable reference. In some embodiments, the term “reduced” or equivalent terms refers to a complete or essentially complete inhibition, i.e., a reduction to zero or essentially to zero. In some embodiments, the term “increased” or “induced” refers to an increase in the level of an assessed value by at least 10%, at least 20%, at least 30%, 21 11979815v1 P-627574-PC at least 40%, at least 50%, at least 80%, at least 100%, at least 200%, at least 500%, or higher, as compared to a comparable reference. [0069] Ionizable: The term “ionizable” refers to a compound or group or atom that is charged at a certain pH. In the context of an ionizable amino lipid, such a lipid or a function group or atom thereof bears a positive charge at a certain pH. In some embodiments, an ionizable amino lipid is positively charged at an acidic pH. In some embodiments, an ionizable amino lipid is predominately neutral at physiological pH values, e.g., in some embodiments about 7.0-7.4, but becomes positively charged at lower pH values. In some embodiments, an ionizable amino lipid may have a pKa within a range of about 5 to about 7. [0070] Isolated: The term “isolated” means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not “isolated”, but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is “isolated”. An isolated nucleic acid or protein can exist in substantially purified form [0071] Nucleic acid/ Polynucleotide: As used herein, the term “nucleic acid” refers to a polymer of at least 10 nucleotides or more. In some embodiments, a nucleic acid is or comprises DNA. In some embodiments, a nucleic acid is or comprises RNA. In some embodiments, a nucleic acid is or comprises peptide nucleic acid (PNA). In some embodiments, a nucleic acid is or comprises a single stranded nucleic acid. In some embodiments, a nucleic acid is or comprises a double- stranded nucleic acid. In some embodiments, a nucleic acid comprises both single and double- stranded portions. In some embodiments, a nucleic acid comprises a backbone that comprises one or more phosphodiester linkages. In some embodiments, a nucleic acid comprises a backbone that comprises both phosphodiester and non-phosphodiester linkages. For example, in some embodiments, a nucleic acid may comprise a backbone that comprises one or more phosphorothioate or 5'-N-phosphoramidite linkages and/or one or more peptide bonds, e.g., as in a “peptide nucleic acid”. In some embodiments, a nucleic acid comprises one or more, or all, natural residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, a nucleic acid comprises on or more, or all, non-natural residues. In some embodiments, a non-natural residue comprises a nucleoside analog (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3 - methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2- aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5 - 22 11979815v1 P-627574-PC propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 6-O-methylguanine, 2-thiocytidine, methylated bases, intercalated bases, and combinations thereof). In some embodiments, a non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) as compared to those in natural residues. In some embodiments, a nucleic acid has a nucleotide sequence that encodes a functional gene product such as an RNA or polypeptide. In some embodiments, a nucleic acid has a nucleotide sequence that comprises one or more introns. In some embodiments, a nucleic acid may be prepared by isolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, e.g., in vivo or in vitro, reproduction in a recombinant cell or system, or chemical synthesis. In some embodiments, a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, or 20,000 or more residues or nucleotides long. [0072] Nucleotide: As used herein, the term “nucleotide” refers to its art-recognized meaning. When a number of nucleotides is used as an indication of size, e.g., of a polynucleotide, a certain number of nucleotides refers to the number of nucleotides on a single strand, e.g., of a polynucleotide. [0073] Patient: As used herein, the term “patient” refers to any organism who is suffering or at risk of a disease or disorder or condition. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and/or humans). In some embodiments, a patient is a human. In some embodiments, a patient is suffering from or susceptible to one or more diseases or disorders or conditions. In some embodiments, a patient displays one or more symptoms of a disease or disorder or condition. In some embodiments, a patient has been diagnosed with one or more diseases or disorders or conditions. In some embodiments, a disease or disorder or condition that is amenable to provided technologies is or includes an HSV infection. In some embodiments, a patient is receiving or has received certain therapy to diagnose and/or to treat a disease, disorder, or condition. In some embodiments, a patient is a patient suffering from or susceptible to an HSV infection. 23 11979815v1 P-627574-PC [0074] Pharmaceutical composition: As used herein, the term “pharmaceutical composition” refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, pharmaceutical compositions may be specially formulated for parenteral administration, for example, by subcutaneous, intramuscular, or intravenous injection as, for example, a sterile solution or suspension formulation. [0075] Pharmaceutically effective amount: The term “pharmaceutically effective amount” or “therapeutically effective amount” refers to the amount which achieves a desired reaction or a desired effect alone or together with further doses. In the case of the treatment of a particular disease, a desired reaction in some embodiments relates to inhibition of the course of the disease. In some embodiments, such inhibition may comprise slowing down the progress of a disease and/or interrupting or reversing the progress of the disease. In some embodiments, a desired reaction in a treatment of a disease may be or comprise delay or prevention of the onset of a disease or a condition. An effective amount of pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) described herein will depend, for example, on a disease or condition to be treated, the severity of such a disease or condition, individual parameters of the patient, including, e.g., age, physiological condition, size and weight, the duration of treatment, the type of an accompanying therapy (if present), the specific route of administration and similar factors. Accordingly, doses of pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) described herein may depend on various of such parameters. In the case that a reaction in a patient is insufficient with an initial dose, higher doses (or effectively higher doses achieved by a different, more localized route of administration) may be used. [0076] Poly(A) sequence: As used herein, the term “poly(A) sequence” or “poly-A tail” refers to an uninterrupted or interrupted sequence of adenylate residues which is typically located at the 3'- end of an RNA molecule. Poly(A) sequences are known to those of skill in the art and may follow the 3’-UTR in the RNAs described herein. An uninterrupted poly(A) sequence is characterized by consecutive adenylate residues. In nature, an uninterrupted poly(A) sequence is typical. RNAs disclosed herein can have a poly(A) sequence attached to the free 3'-end of the RNA by a template- independent RNA polymerase after transcription or a poly(A) sequence encoded by DNA and transcribed by a template-dependent RNA polymerase. 24 11979815v1 P-627574-PC [0077] Prevent: As used herein, the term “prevent” or “prevention” when used in connection with the occurrence of a disease, disorder, and/or condition, refers to reducing the risk of developing the disease, disorder and/or condition and/or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time. [0078] Recombinant: The term “recombinant” in the context of the present disclosure means “made through genetic engineering”. In some embodiments, a “recombinant” entity such as a recombinant nucleic acid in the context of the present disclosure is not naturally occurring. [0079] Reference: As used herein, the term “reference” describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value. In some embodiments, a reference or control is tested and/or determined substantially simultaneously with the testing or determination of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as would be understood by those skilled in the art, a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment. Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and/or comparison to a particular possible reference or control. [0080] Risk: As will be understood from context, “risk” of a disease, disorder, and/or condition refers to a likelihood that a particular individual will develop the disease, disorder, and/or condition. In some embodiments, risk is expressed as a percentage. In some embodiments, risk is from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90 up to 100%. In some embodiments risk is expressed as a risk relative to a risk associated with a reference sample or group of reference samples. In some embodiments, a reference sample or group of reference samples have a known risk of a disease, disorder, condition and/or event. In some embodiments a reference sample or group of reference samples are from individuals comparable to a particular individual. In some embodiments, relative risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. In some embodiments, risk may reflect one or more genetic attributes, e.g., which may predispose an individual toward development (or not) of a particular disease, disorder and/or condition. In some embodiments, risk may reflect one or more epigenetic events or attributes and/or one or more lifestyle or environmental events or attributes. 25 11979815v1 P-627574-PC [0081] Selective or specific: The term “selective” or “specific”, when used herein in reference to an agent having an activity, is understood by those skilled in the art to mean that the agent discriminates between potential target entities, states, or cells. For example, in some embodiments, an agent is said to bind “specifically” to its target if it binds preferentially with that target in the presence of one or more competing alternative targets. In many embodiments, specific interaction is dependent upon the presence of a particular structural feature of the target entity (e.g., an epitope, a cleft, a binding site). It is to be understood that specificity need not be absolute. In some embodiments, specificity may be evaluated relative to that of a target-binding moiety for one or more other potential target entities (e.g., competitors). In some embodiments, specificity is evaluated relative to that of a reference specific binding moiety. In some embodiments, specificity is evaluated relative to that of a reference non-specific binding moiety. [0082] Subject: As used herein, the term “subject” refers to an organism to be administered with a composition described herein, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, domestic pets, etc.) and humans. In some embodiments, a subject is a human subject. In some embodiments, a subject is suffering from a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject is susceptible to a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject displays one or more symptoms or characteristics of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject displays one or more non-specific symptoms of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject does not display any symptom or characteristic of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition (e.g., an HSV infection). In some embodiments, a subject is a patient. In some embodiments, a subject is an individual to whom diagnosis and/or therapy is and/or has been administered. [0083] Suffering from: An individual who is “suffering from” a disease, disorder, and/or condition has been diagnosed with and/or displays one or more symptoms of a disease, disorder, and/or condition. [0084] Susceptible to: An individual who is “susceptible to” a disease, disorder, and/or condition is one who has a higher risk of developing the disease, disorder, and/or condition than does a member of the general public. In some embodiments, an individual who is susceptible to a disease, 26 11979815v1 P-627574-PC disorder and/or condition may not have been diagnosed with the disease, disorder, and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition may exhibit symptoms of the disease, disorder, and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition may not exhibit symptoms of the disease, disorder, and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition will develop the disease, disorder, and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition will not develop the disease, disorder, and/or condition. [0085] Synthetic: As used herein, the term “synthetic” refers to an entity that is artificial, or that is made with human intervention, or that results from synthesis rather than naturally occurring. For example, in some embodiments, a synthetic nucleic acid or polynucleotide refers to a nucleic acid molecule that is chemically synthesized, e.g., in some embodiments by solid-phase synthesis. In some embodiments, the term “synthetic” refers to an entity that is made outside of biological cells. For example, in some embodiments, a synthetic nucleic acid or polynucleotide refers to a nucleic acid molecule (e.g., an RNA) that is produced by in vitro transcription using a template. [0086] Therapy: The term “therapy” refers to an administration or delivery of an agent or intervention that has a therapeutic effect and/or elicits a desired biological and/or pharmacological effect (e.g., has been demonstrated to be statistically likely to have such effect when administered to a relevant population). In some embodiments, a therapeutic agent or therapy is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition. In some embodiments, a therapeutic agent or therapy is a medical intervention (e.g., surgery, radiation, phototherapy) that can be performed to alleviate, relieve, inhibit, present, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition. [0087] Three prime untranslated region: As used herein, the terms “three prime untranslated region” or “3' UTR” refer to a sequence of an RNA molecule that begins following a stop codon of a coding region of an open reading frame sequence. In some embodiments, the 3' UTR begins immediately after a stop codon of a coding region of an open reading frame sequence, e.g., in its natural context. In other embodiments, the 3' UTR does not begin immediately after stop codon of the coding region of an open reading frame sequence, e.g., in its natural context. 27 11979815v1 P-627574-PC [0088] Threshold level (e.g., acceptance criteria): As used herein, the term “threshold level” refers to a level that are used as a reference to attain information on and/or classify the results of a measurement, for example, the results of a measurement attained in an assay. For example, in some embodiments, a threshold level means a value measured in an assay that defines the dividing line between two subsets of a population (e.g. a batch that satisfy quality control criteria vs. a batch that does not satisfy quality control criteria). Thus, a value that is equal to or higher than the threshold level defines one subset of the population, and a value that is lower than the threshold level defines the other subset of the population. A threshold level can be determined based on one or more control samples or across a population of control samples. A threshold level can be determined prior to, concurrently with, or after the measurement of interest is taken. In some embodiments, a threshold level can be a range of values. [0089] Treat: As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition. In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition, for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition. In some embodiments, treatment may be administered to a subject at a later-stage of disease, disorder, and/or condition. [0090] Vaccination: As used herein, the term “vaccination” refers to the administration of a composition intended to generate an immune response, for example to a disease-associated (e.g., disease-causing) agent. In some embodiments, vaccination can be administered before, during, and/or after exposure to a disease-associated agent, and in certain embodiments, before, during, and/or shortly after exposure to the agent. In some embodiments, vaccination includes multiple administrations, appropriately spaced in time, of a vaccine composition. In some embodiments, vaccination generates an immune response to an infectious agent. [0091] Vaccine: As used herein, the term “vaccine” refers to a composition that induces an immune response upon administration to a subject. In some embodiments, an induced immune response provides protective immunity. [0092] Variant: As used herein in the context of molecules, e.g., nucleic acids, proteins, or small molecules, the term “variant” refers to a molecule that shows significant structural identity with a 28 11979815v1 P-627574-PC reference molecule but differs structurally from the reference molecule, e.g., in the presence or absence or in the level of one or more chemical moieties as compared to the reference entity. In some embodiments, a variant also differs functionally from its reference molecule. In general, whether a particular molecule is properly considered to be a “variant” of a reference molecule is based on its degree of structural identity with the reference molecule. As will be appreciated by those skilled in the art, any biological or chemical reference molecule has certain characteristic structural elements. A variant, by definition, is a distinct molecule that shares one or more such characteristic structural elements but differs in at least one aspect from the reference molecule. In some embodiments, a variant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in amino acid or nucleotide sequence and/or one or more differences in chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nucleic acid (e.g., that are attached to the polypeptide or nucleic acid backbone). In some embodiments, a variant polypeptide or nucleic acid shows an overall sequence identity with a reference polypeptide or nucleic acid that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 99%. In some embodiments, a variant polypeptide or nucleic acid does not share at least one characteristic sequence element with a reference polypeptide or nucleic acid. In some embodiments, a reference polypeptide or nucleic acid has one or more biological activities. In some embodiments, a variant polypeptide or nucleic acid shares one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid lacks one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid shows a reduced level of one or more biological activities as compared to the reference polypeptide or nucleic acid. In some embodiments, a polypeptide or nucleic acid of interest is considered to be a “variant” of a reference polypeptide or nucleic acid if it has an amino acid or nucleotide sequence that is identical to that of the reference but for a small number of sequence alterations at particular positions. Typically, fewer than about 20%, about 15%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, or about 2% of the residues in a variant are substituted, inserted, or deleted, as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, or about 1 substituted residues as compared to a reference. Often, a variant polypeptide or nucleic acid comprises a very small number (e.g., fewer than about 5, about 4, about 3, about 2, or about 1) 29 11979815v1 P-627574-PC number of substituted, inserted, or deleted, functional residues (i.e., residues that participate in a particular biological activity) relative to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises not more than about 5, about 4, about 3, about 2, or about 1 addition or deletion, and, in some embodiments, comprises no additions or deletions, as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises fewer than about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 10, about 9, about 8, about 7, about 6, and commonly fewer than about 5, about 4, about 3, or about 2 additions or deletions as compared to the reference. In some embodiments, a reference polypeptide or nucleic acid is one found in nature. [0093] Vector: as used herein, refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a “plasmid”, which refers to a circular double stranded DNA loop into which additional DNA segments may be ligated. Another type of vector is a viral vector, wherein additional DNA segments may be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as “expression vectors.” In some embodiments, known techniques may be used, for example, for generation or manipulation of recombinant DNA, for oligonucleotide synthesis, and for tissue culture and transformation (e.g., electroporation, lipofection). Enzymatic reactions and purification techniques may be performed according to manufacturer's specifications or as commonly accomplished in the art or as described herein. The foregoing techniques and procedures may be generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification. See e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual (4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2012)), which is incorporated herein by reference for any purpose. [0094] All literature and similar material cited in this application, including, but not limited to, patents, patent applications, articles, books, treatises, and web pages, regardless of the format of such literature and similar materials, are expressly incorporated by reference in their entirety. In the event that one or more of the incorporated literature and similar materials differs from or 30 11979815v1 P-627574-PC contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like, this application controls. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described in any way. DETAILED DESCRIPTION Compositions [0095] In some embodiments, the present disclosure provides compositions comprising one or more RNAs (also referred to herein as “polyribonucleotides”). In some embodiments, one or more RNAs encodes a Herpes Simplex Virus (HSV) glycoprotein or immunogenic fragment thereof. In some embodiments, an RNA is a modified RNA as described herein below. [0096] In some embodiments, an immunogenic fragment of an HSV glycoprotein comprises the ectodomain of the glycoprotein, or portion thereof. In another embodiment, an immunogenic fragment consists of the ectodomain of the glycoprotein, or portion thereof. [0097] In some embodiments, the present disclosure provides a composition comprising one or more nucleoside modified RNAs. In some embodiments, a modified RNA encodes a Herpes Simplex Virus (HSV) glycoprotein or immunogenic fragment thereof. In some embodiments, modified RNA comprises one or more pseudouridine or pseudouridine family residues. [0098] In some embodiments, an HSV glycoprotein comprises glycoprotein D (gD), glycoprotein C (gC), glycoprotein E (gE), glycoprotein B (gB), glycoprotein H (gH), glycoprotein L (gL), glycoprotein I (gI), or any combinations thereof. [0099] In some embodiments, the present disclosure provides a composition comprising one or more modified RNAs encoding HSV gD, gC, gE, gB, gH, gL, gI, or immunogenic fragments thereof. In some embodiments, the modified RNAs comprise pseudouridine-modified RNAs. [0100] In some embodiments, the present disclosure provides compositions comprising an RNA encoding HSV gD or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV gC or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV gE or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV gB or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV gH or an immunogenic fragment thereof. In 31 11979815v1 P-627574-PC another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV gL or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV gI or an immunogenic fragment thereof. [0101] In some embodiments, the present disclosure provides a composition comprising: (a) an RNA encoding HSV gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV gC or an immunogenic fragment thereof. [0102] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV gE or an immunogenic fragment thereof. [0103] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV gC or an immunogenic fragment thereof; and (b) an RNA encoding HSV gE or an immunogenic fragment thereof. [0104] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV gD or an immunogenic fragment thereof; (b) an RNA encoding HSV gC or an immunogenic fragment thereof, and (c) an RNA encoding HSV gE or an immunogenic fragment thereof. [0105] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV gD or an immunogenic fragment thereof; (b) an RNA encoding HSV gC or an immunogenic fragment thereof, (c) an RNA encoding HSV gE or an immunogenic fragment thereof; and (d) an RNA encoding HSV gB or an immunogenic fragment thereof. [0106] In some embodiments, the HSV glycoproteins are HSV-2 glycoproteins or immunogenic fragments thereof. In another embodiment, the HSV glycoproteins are HSV-1 glycoproteins or immunogenic fragments thereof. In some embodiments, the HSV glycoproteins comprise both HSV-2 glycoproteins and HSV-1 glycoproteins or immunogenic fragments thereof. In another embodiment, the HSV glycoproteins comprise a mixture of HSV-2 glycoproteins and HSV-1 glycoproteins or immunogenic fragments thereof. [0107] In some embodiments, the present disclosure provides compositions comprising an RNA encoding HSV-2 gD or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-2 gC or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-2 gE or an immunogenic fragment thereof. In another 32 11979815v1 P-627574-PC embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-2 gE or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-2 gB or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-2 gH or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-2 gL or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-2 gI or an immunogenic fragment thereof. [0108] In some embodiments, the present disclosure provides a composition comprising: (a) an RNA encoding HSV-2 gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 2 gC or an immunogenic fragment thereof. [0109] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV-2 gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 2 gE or an immunogenic fragment thereof. [0110] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV-2 gC or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 2 gE or an immunogenic fragment thereof. [0111] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV-2 gD or an immunogenic fragment thereof; (b) an RNA encoding HSV-2 gC or an immunogenic fragment thereof, and (c) an RNA encoding HSV-2 gE or an immunogenic fragment thereof. [0112] In some embodiments, the present disclosure provides compositions comprising an RNA encoding HSV-1 gD or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gC or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gE or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gE or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gB or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gH or an immunogenic fragment thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gL or an immunogenic fragment 33 11979815v1 P-627574-PC thereof. In another embodiment, the present disclosure provides compositions comprising an RNA encoding HSV-1 gI or an immunogenic fragment thereof. [0113] In some embodiments, the present disclosure provides a composition comprising: (a) an RNA encoding HSV-1 gD or fragment thereof; and (b) an RNA encoding HSV-1 gC or fragment thereof. [0114] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV-1 gD or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 1 gE or an immunogenic fragment thereof. [0115] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV-1 gC or an immunogenic fragment thereof; and (b) an RNA encoding HSV- 1 gE or an immunogenic fragment thereof. [0116] In another embodiment, the present disclosure provides a composition comprising: (a) an RNA encoding HSV-1 gD or an immunogenic fragment thereof; (b) an RNA encoding HSV-1 gC or an immunogenic fragment thereof, and (c) an RNA encoding HSV-1 gE or an immunogenic fragment thereof. [0117] In some embodiments, any of the compositions as described herein consists essentially of one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof. In another embodiment, any of the compositions as described herein consists of one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof. [0118] In another embodiment, the present disclosure provides compositions comprising an RNA encoding an HSV gD protein or immunogenic fragment thereof, an RNA encoding an HSV gC protein or immunogenic fragment thereof, an RNA encoding an HSV gE protein or immunogenic fragment thereof, and RNAs encoding one or more additional HSV glycoproteins. In some embodiments, said additional HSV glycoproteins comprise gB or immunogenic fragment thereof, gH or immunogenic fragment thereof, gL or immunogenic fragment thereof, gI or immunogenic fragment thereof, or any combination thereof. In some embodiments, said additional HSV glycoproteins comprise glycoprotein M (gM), glycoprotein N (gN), glycoprotein K (gK), glycoprotein G (gG), glycoprotein J (gJ), or an immunogenic fragment thereof. [0119] In some embodiments, compositions of the present disclosure and for use in the methods of the present disclosure comprise both HSV-2 glycoproteins or glycoprotein fragments and HSV- 1 glycoproteins or glycoprotein fragments. In another embodiment, compositions of the present 34 11979815v1 P-627574-PC disclosure and for use in the methods of the present disclosure comprise a mixture of HSV-2 glycoproteins or glycoprotein fragments and HSV-1 glycoproteins or glycoprotein fragments. For example, in some embodiments, a composition of the present disclosure comprises HSV-2 gC, HSV-1 gD, and HSV-2 gE, or immunogenic fragments thereof. In another embodiment, a composition of the present disclosure comprises HSV-1 gC, HSV-2 gD, and HSV-2 gE, or immunogenic fragments thereof. In another embodiment, a composition of the present disclosure comprises HSV-2 gC, HSV-2 gD, and HSV-1 gE, or immunogenic fragments thereof. In another embodiment, a composition of the present disclosure comprises HSV-1 gC, HSV-1 gD, and HSV- 2 gE, or immunogenic fragments thereof. In another embodiment, a composition of the present disclosure comprises HSV-1 gC, HSV-2 gD, and HSV-1 gE, or immunogenic fragments thereof. In another embodiment, a composition of the present disclosure comprises HSV-2 gC, HSV-1 gD, and HSV-1 gE, or immunogenic fragments thereof. [0120] In another embodiment, the compositions of the present disclosure comprise one or more additional (i) HSV-1 glycoproteins or immunogenic fragments thereof, (ii) HSV-2 glycoproteins or immunogenic fragments thereof, or (iii) both HSV-1 or immunogenic fragments thereof and HSV-2 glycoproteins or immunogenic fragments thereof, as described herein. For example, in some embodiments, a composition of the present disclosure comprising HSV-2 gC, HSV-1 gD, and HSV-2 gE may further comprise HSV-1 gI. In another embodiment, a composition of the present disclosure comprising HSV-2 gC, HSV-2 gD, and HSV-2 gE may further comprise HSV- 1 gB. Each of the possible combinations of HSV-1 and HSV-2 glycoproteins represents a separate embodiment of the disclosure. [0121] As used herein, “encoding” refers to an RNA molecule that contains a gene that encodes a protein of interest or a fragment thereof. In another embodiment, an RNA molecule comprises a coding sequence that encodes a protein of interest or a fragment thereof. In another embodiment, one or more other proteins or fragments thereof is also encoded. In another embodiment, the protein of interest or a fragment thereof is the only protein encoded. Each possibility represents a separate embodiment of the present disclosure. [0122] “Immunogenic fragment” refers to a portion of a protein that is immunogenic and elicits a protective immune response when administered to a subject. [0123] “Immunogenicity” or “immunogenic” is used herein to refer to the innate ability of a protein, peptide, protein fragment, nucleic acid, antigen or organism to elicit an immune response in an animal when the protein, peptide, nucleic acid, antigen or organism is administered to the 35 11979815v1 P-627574-PC animal. Thus, “enhancing the immunogenicity” refers to increasing the ability of a protein, peptide, protein fragment, nucleic acid, antigen or organism to elicit an immune response in an animal when the protein, peptide, protein fragment, nucleic acid, antigen or organism is administered to an animal. The increased ability of a protein, peptide, protein fragment, nucleic acid, antigen or organism to elicit an immune response can be measured by, in some embodiments, a greater number of antibodies to a protein, peptide, protein fragment, nucleic acid, antigen or organism, a greater diversity of antibodies to an antigen or organism, a greater number of T-cells specific for a protein, peptide, protein fragment, nucleic acid, antigen or organism, a greater cytotoxic or helper T-cell response to a protein, peptide, protein fragment, nucleic acid, antigen or organism, and the like. [0124] In some embodiments, a protein, peptide, protein fragment, nucleic acid or organism can be antigenic. “Antigenic” refers to a protein, peptide, protein fragment, nucleic acid, or organism capable of specifically interacting with an antigen recognition molecule of the immune system, e.g., an immunoglobulin (antibody) or T cell antigen receptor. An antigenic protein, peptide, protein fragment contains, in another embodiment, an epitope of at least about 8 amino acids (AA). An antigenic portion of a protein, peptide, protein fragment, nucleic acid, or organism, also called herein an “epitope” can be a portion that is immunodominant for antibody or T cell receptor recognition, or it can be a portion used to generate an antibody to the molecule by conjugating an antigenic portion to a carrier (e.g., a carrier polypeptide) for immunization. A molecule that is antigenic need not be itself immunogenic, i.e., capable of eliciting an immune response without a carrier. [0125] “Functional” is used herein to refer to the innate ability of a protein, peptide, nucleic acid, fragment or a variant thereof to exhibit a biological activity or function. In some embodiments, such a biological function is its binding property to an interaction partner, e.g., a membrane- associated receptor, and in another embodiment, its trimerization property. In the case of functional fragments and the functional variants of the disclosure, these biological functions may in fact be changed, e.g., with respect to their specificity or selectivity, but with retention of the basic biological function. [0126] In some embodiments, the term “fragment” is used herein to refer to a protein or polypeptide that is shorter or comprises fewer amino acids than the full length protein or polypeptide. In another embodiment, fragment refers to a nucleic acid encoding the protein fragment that is shorter or comprises fewer nucleotides than the full length nucleic acid. In another 36 11979815v1 P-627574-PC embodiment, the fragment is an N-terminal fragment. In another embodiment, the fragment is a C-terminal fragment. In some embodiments, the fragment is an intrasequential section of the protein, peptide, or nucleic acid. In another embodiment, the fragment is an immunogenic intrasequential section of the protein, peptide or nucleic acid. In another embodiment, the fragment is a functional intrasequential section within the protein, peptide or nucleic acid. In another embodiment, the fragment is an N-terminal immunogenic fragment. In some embodiments, the fragment is a C-terminal immunogenic fragment. In another embodiment, the fragment is an N- terminal functional fragment. In another embodiment, the fragment is a C-terminal functional fragment. In another embodiment, the fragment contains pieces of the protein linked together or pieces of multiple proteins linked together. In some embodiments, a fragment is a domain (e.g., an ectodomain). [0127] Thus, in some embodiments, an “immunogenic fragment” of a protein as described in the present disclosure refers to a portion of the protein that is immunogenic, in one embodiment and in another embodiment, elicits a protective immune response when administered to a subject. [0128] In another aspect, the present disclosure provides compositions comprising RNAs, wherein each of said RNAs encodes a) HSV glycoprotein D (gD) or an immunogenic fragment thereof, b) HSV glycoprotein C (gC) or an immunogenic fragment thereof, c) HSV glycoprotein E (gE) or an immunogenic fragment thereof, or any combination thereof. [0129] In some embodiments, the present disclosure provides a composition comprising an RNA encoding an HSV gD or an immunogenic fragment thereof, an RNA encoding an HSV gC or an immunogenic fragment thereof, and an RNA encoding an HSV gE or an immunogenic fragment thereof. [0130] In some embodiments, compositions of RNA encoding HSV-1 gD (gD1) or an immunogenic fragment thereof are protective against HSV-1 infection. In some embodiments, combination compositions of RNA encoding HSV gC (gC1) or an immunogenic fragment thereof /HSV-1 gD (gD1) or an immunogenic fragment thereof /HSV-1 gE (gE1) or an immunogenic fragment thereof can are protective against HSV-1. Further, as provided herein, compositions of RNA encoding gD2 or an immunogenic fragment thereof can be protective against HSV-2 infection (Figures 7-10). Further, combination compositions of RNA encoding gC2 or an immunogenic fragment thereof /gD2 or an immunogenic fragment thereof /gE2 or an immunogenic fragment thereof can confer superior protection compared with compositions containing RNA encoding gC2 alone, gD2 alone, or gE2 alone. 37 11979815v1 P-627574-PC [0131] In some embodiments, inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure can increase the efficaciousness of anti-gD antibodies elicited by the composition. In some embodiments, inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure can increase the dose of RNA encoding gD or an immunogenic fragment thereof required to elicit antibodies that inhibit binding of gD to a cellular receptor. In another embodiment, inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure can decrease the dose of RNA encoding gD or an immunogenic fragment thereof required to elicit antibodies that inhibit binding of gD or an immunogenic fragment thereof to a cellular receptor when a dose of RNA encoding a gD or an immunogenic fragment thereof is administered separately from RNAs encoding the gC protein, gE protein or immunogenic fragments thereof. [0132] In another embodiment, inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure enhances the effectiveness of an innate immune response. In another embodiment, the innate immune response is an antibody-mediated immune response. In another embodiment, the innate immune response is a non-antibody-mediated immune response. In another embodiment, the innate immune response is an NK (natural killer) cell response. In another embodiment, the innate immune response is any other innate immune response known in the art. [0133] In another embodiment, inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure increases the efficaciousness of antibodies elicited by the composition against one of the above glycoproteins. In another embodiment, inclusion of an RNA encoding gC or an immunogenic fragment thereof, and/or an RNA encoding gE or an immunogenic fragment thereof in the composition of the present disclosure decreases the dose of one of the above glycoproteins required to elicit antibodies that inhibit binding of the glycoprotein to a cellular receptor thereof, when a dose of one of the glycoproteins is administered separately from one of the other glycoproteins. 38 11979815v1 P-627574-PC [0134] In some embodiments, a composition comprises (i) one or more RNAs encoding HSV glycoproteins or immunogenic fragments thereof and (ii) lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes. Glycoprotein D HSV-1 gD [0135] In some embodiments, a composition of the present disclosure comprises an RNA encoding HSV-1 gD protein. In some embodiments, a composition comprises an RNA encoding a fragment of an HSV-1 gD protein (e.g., an immunogenic fragment). [0136] In some embodiments, a nucleotide sequence of the RNA encoding an HSV-1 gD fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCU GCUGCUGCUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCAAGUACGC CCUGGCCGACGCCUCCCUGAAGAUGGCCGACCCCAACCGCUUCCGCGGCAAGG ACCUGCCCGUGCUGGACCAGCUGACCGACCCCCCCGGCGUGCGCCGCGUGUAC CACAUCCAGGCCGGCCUGCCCGACCCCUUCCAGCCCCCCUCCCUGCCCAUCACC GUGUACUACGCCGUGCUGGAGCGCGCCUGCCGCUCCGUGCUGCUGAACGCCCC CUCCGAGGCCCCCCAGAUCGUGCGCGGCGCCUCCGAGGACGUGCGCAAGCAGC CCUACAACCUGACCAUCGCCUGGUUCCGCAUGGGCGGCAACUGCGCCAUCCCC AUCACCGUGAUGGAGUACACCGAGUGCUCCUACAACAAGUCCCUGGGCGCCUG CCCCAUCCGCACCCAGCCCCGCUGGAACUACUACGACUCCUUCUCCGCCGUGU CCGAGGACAACCUGGGCUUCCUGAUGCACGCCCCCGCCUUCGAGACCGCCGGC ACCUACCUGCGCCUGGUGAAGAUCAACGACUGGACCGAGAUCACCCAGUUCAU CCUGGAGCACCGCGCCAAGGGCUCCUGCAAGUACGCCCUGCCCCUGCGCAUCC CCCCCUCCGCCUGCCUGUCCCCCCAGGCCUACCAGCAGGGCGUGACCGUGGAC UCCAUCGGCAUGCUGCCCCGCUUCAUCCCCGAGAACCAGCGCACCGUGGCCGU GUACUCCCUGAAGAUCGCCGGCUGGCACGGCCCCAAGGCCCCCUACACCUCCA CCCUGCUGCCCCCCGAGCUGUCCGAGACCCCCAACGCCACCCAGCCCGAGCUGG CCCCCGAGGACCCCGAGGACUCCGCCCUGCUGGAGGACCCCGUGGGCACCGUG GCCCCCCAGAUCCCCCCCAACUGGCACAUCCCCUCCAUCCAGGACGCCGCCACC CCCUACUAACUAGUAGUGACUGACUAGGAUCUGGUUACCACUAAACCAGCCUCAA 39 11979815v1 P-627574-PC GAACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUGU UGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUC ACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAC (SEQ ID NO: 1) [0137] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 19) to assist expression of the HSV-1 gD fragment. In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264). [0138] In another embodiment, a nucleotide sequence of the RNA encoding an HSV-1 gD fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or any combinations thereof. In some embodiments, the sequence of the HSV-1 gD fragment is as set forth in SEQ ID NO: 22. [0139] In some embodiments, an HSV-1 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-331 of gD (e.g., from HSV- 1 Patton strain), as set forth in the following amino acid sequence: KYALADASLKMADPNRFRGKDLPVLDQLTDPPGVRRVYHIQAGLPDPFQPPSLPITV YYAVLERACRSVLLNAPSEAPQIVRGASEDVRKQPYNLTIAWFRMGGNCAIPITVME YTECSYNKSLGACPIRTQPRWNYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKIN DWTEITQFILEHRAKGSCKYALPLRIPPSACLSPQAYQQGVTVDSIGMLPRFIPENQRT VAVYSLKIAGWHGPKAPYTSTLLPPELSETPNATQPELAPEDPEDSALLEDPVGTVAP QIPPNWHIPSIQDAATPY (SEQ ID NO: 2 ) [0140] In some embodiments, an HSV-1 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 2. In some embodiments, an HSV-1 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 2. [0141] In some embodiments, the HSV-1 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-333 of gD (e.g., from HSV- 1 Patton strain), as set forth in the following amino acid sequence: KYALADASLKMADPNRFRGKDLPVLDQLTDPPGVRRVYHIQAGLPDPFQPPSLPITV YYAVLERACRSVLLNAPSEAPQIVRGASEDVRKQPYNLTIAWFRMGGNCAIPITVME 40 11979815v1 P-627574-PC YTECSYNKSLGACPIRTQPRWNYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKIN DWTEITQFILEHRAKGSCKYALPLRIPPSACLSPQAYQQGVTVDSIGMLPRFIPENQRT VAVYSLKIAGWHGPKAPYTSTLLPPELSETPNATQPELAPEDPEDSALLEDPVGTVAP QIPPNWHIPSIQDAATPYHP (SEQ ID NO: 134) [0142] In some embodiments, an HSV-1 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 134. In some embodiments, an HSV-1 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 134. [0143] In some embodiments, full length HSV-1 gD encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MGGAAARLGAVILFVVIVGLHGVRGKYALADASLKLADPNRFRRKDLPVLDQLTDP PGVRRVYHIQAGLPDPFQPPSLPITVYYAVLERACRSVLLNAPSEAPQIVRGASEDVR KQPYNLTIAWFRMGGNCAIPITVMEYTECSYNKSLGACPIRTQPRWNYYDSFSAVSE DNLGFLMHAPAFETAGTYLRLVKINDWTEITQFILEHRAKGSCKYALPLRIPPSACLS PQAYQQGVTVDSIGMLPRFIPENQRTVAVYSLKIAGWHGPKAPYTSTLLPPELSETPN ATQPELAPEAPEDSALLEDPVGTVAPQIPPNWHIPSIQDAATPYHPPATPNNMGLIAG AVGGSLLAALVICGIVYWMRRRTQKAPKRIRLPHIREDDQPSSHQPLFY (SEQ ID NO: 3) [0144] In some embodiments, an HSV-1 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 3. In some embodiments, an HSV-1 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 3. [0145] In another embodiment, an HSV-1 gD, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any one of the following GenBank Accession Numbers: AAL90884.1 (KHS2 strain), AAL90883.1 (KHS1 strain), AAK93950.1 (F strain), AAB59754.1 (F strain), AAA19631.1 (mutant strain not identified), AAA19630.1 (mutant strain not identified), or AAA19629.1 (strain not identified). [0146] In another embodiment, an HSV-1 gD, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid 41 11979815v1 P-627574-PC sequences as set forth in any of the following GenBank Accession Numbers: A1Z0Q5.2, AAA45780.1, AAA45785.1, AAA45786.1, AAA96682.1, AAK19597.1, AAN74642.1, ABI63524.1, ABM52978.1, ABM52979.1, ABM52980.1, ABM52981.1, ABM66847.1, ABM66848.1, ACM62295.1, ADD60053.1, ADD60130.1, ADM22389.1, ADM22466.1, ADM22542.1, ADM22619.1, ADM22696.1, ADM22773.1, ADM22849.1, ADM22926.1, ADM23003.1, ADM23079.1, ADM23155.1, ADM23231.1, ADM23309.1, ADM23383.1, ADM23457.1, ADM23531.1, ADM23605.1, ADM23680.1, ADM23755.1, ADM23831.1, AEQ77097.1, AER37647.1, AER37715.1, AER37786.1, AER37857.1, AER37929.1, AER38000.1, AER38070.1, AFE62894.1, AFH41180.1, AFI23657.1, AFK50415.1, AFP86430.1, AGZ01928.1, AIR95858.1, AJE60009.1, AJE60080.1, AJE60151.1, AJE60222.1, AJE60293.1, AJE60439.1, AKE48645.1, AKG59246.1, AKG59318.1, AKG59391.1, AKG59462.1, AKG59536.1, AKG59609.1, AKG59682.1, AKG59755.1, AKG59826.1, AKG59898.1, AKG59972.1, AKG60046.1, AKG60118.1, AKG60189.1, AKG60261.1, AKG60334.1, AKG60404.1, AKG60474.1, AKG60546.1, AKG60620.1, AKG60692.1, AKG60763.1, AKG60835.1, AKG60906.1, AKG60978.1, AKG61050.1, AKG61123.1, AKG61194.1, AKG61267.1, AKG61339.1, AKG61411.1, AKG61484.1, AKG61556.1, AKG61629.1, AKG61703.1, AKG61774.1, AKG61847.1, AKG61920.1, AKG61993.1, AKH80463.1, AKH80536.1, ALM22635.1, ALM22709.1, ALM22783.1, ALM22857.1, ALO18662.1, ALO18738.1, AMB65662.1, AMB65735.1, AMB65809.1, AMB65885.1, AMB65956.1, AMN09832.1, ANN83964.1, ANN84041.1, ANN84117.1, ANN84194.1, ANN84271.1, ANN84348.1, ANN84424.1, ANN84500.1, ANN84577.1, ANN84653.1, ANN84730.1, ANN84806.1, ANN84883.1, ANN84959.1, ANN85036.1, ANN85112.1, ANN85187.1, ANN85264.1, ANN85341.1, ANN85416.1, ANN85494.1, ANN85571.1, ANN85648.1, ANN85724.1, ANN85801.1, AOY34093.1, AOY34141.1, AOY34243.1, AOY34271.1, AOY34337.1, AOY36685.1, ARB08957.1, ARO37961.1, ARO37962.1, ARO37963.1, ARO37964.1, ARO37965.1, ARO37966.1, ARO37967.1, ARO37968.1, ARO37969.1, ARO37970.1, ARO37971.1, ARO37972.1, ARO37973.1, ARO37974.1, ARO37975.1, ARO37976.1, ARO37977.1, ARO37978.1, ARO37979.1, ARO37980.1, ARO37981.1, ARO37982.1, ARO37983.1, ARO37984.1, ARO37985.1, ARO37986.1, ARO37987.1, ARO37988.1, ARO37989.1, ARO37990.1, ARO37991.1, ARO37992.1, ARO37993.1, ARO37994.1, ARO37995.1, ARO37996.1, ARO37997.1, ARO37998.1, ARO37999.1, ASM47664.1, ASM47741.1, ASM47818.1, ASM47893.1, BAM73419.1, CAA26060.1, 42 11979815v1 P-627574-PC CAA32283.1, CAA32284.1, CAA32289.1, CAA38245.1, CAT05431.1, P06476.1, P36318.1, P57083.1, P68331.1, Q05059.1, Q69091.1, SBO07792.1, SBO07819.1, SBO07855.1, SBO07869.1, SBO07887.1, SBO07908.1, SBS69553.1, SBS69561.1, SBS69579.1, SBS69625.1, SBS69688.1, SBS69694.1, SBS69717.1, SBS69727.1, SBS69811.1, SBT69395.1, SCL76902.1, VGBEDZ, or YP_009137141.1. HSV-2 gD [0147] In another embodiment, a composition comprises an RNA encoding an HSV-2 gD protein. In another embodiment, a composition comprises an RNA encoding a fragment of an HSV-2 gD protein (e.g., an immunogenic fragment). [0148] In some embodiments, a nucleotide sequence of the RNA encoding an HSV-2 gD fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAA GCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAGCAAU UUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGACCCGCCUGACCGUGCU GGCCCUGCUGGCCGGCCUGCUGGCCUCCUCCCGCGCCAAGUACGCCCUGGCCG ACCCCUCCCUGAAGAUGGCCGACCCCAACCGCUUCCGCGGCAAGAACCUGCCCGU GCUGGACCAGCUGACCGACCCCCCCGGCGUGAAGCGCGUGUACCACAUCCAGCCC UCCCUGGAGGACCCCUUCCAGCCCCCCUCCAUCCCCAUCACCGUGUACUACGCCGU GCUGGAGCGCGCCUGCCGCUCCGUGCUGCUGCACGCCCCCUCCGAGGCCCCCCAGA UCGUGCGCGGCGCCUCCGACGAGGCCCGCAAGCACACCUACAACCUGACCAUCGC CUGGUACCGCAUGGGCGACAACUGCGCCAUCCCCAUCACCGUGAUGGAGUACACC GAGUGCCCCUACAACAAGUCCCUGGGCGUGUGCCCCAUCCGCACCCAGCCCCGCU GGUCCUACUACGACUCCUUCUCCGCCGUGUCCGAGGACAACCUGGGCUUCCUGAU GCACGCCCCCGCCUUCGAGACCGCCGGCACCUACCUGCGCCUGGUGAAGAUCAAC GACUGGACCGAGAUCACCCAGUUCAUCCUGGAGCACCGCGCCCGCGCCUCCUGCA AGUACGCCCUGCCCCUGCGCAUCCCCCCCGCCGCCUGCCUGACCUCCAAGGCCUAC CAGCAGGGCGUGACCGUGGACUCCAUCGGCAUGCUGCCCCGCUUCAUCCCCGAGA ACCAGCGCACCGUGGCCCUGUACUCCCUGAAGAUCGCCGGCUGGCACGGCCCCAA GCCCCCCUACACCUCCACCCUGCUGCCCCCCGAGCUGUCCGACACCACCAACGCCA CCCAGCCCGAGCUGGUGCCCGAGGACCCCGAGGACUCCGCCCUGCUGGAGGACCC CGCCGGCACCGUGUCCUCCCAGAUCCCCCCCAACUGGCACAUCCCCUCCAUCCAGG ACGUGGCCCCCCACCACUAACUAGUAGUGACUGACUAGGAUCUGGUUACCACUAAAC 43 11979815v1 P-627574-PC CAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUAC AAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUU CUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAC (SEQ ID NO: 4) [0149] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequence (SEQ ID NO: 243). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 20) to assist expression of the HSV-2 gD fragment. In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264). [0150] In another embodiment, a nucleotide sequence of the RNA encoding an HSV-2 gD fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or any combinations thereof. In some embodiments, the sequence of the HSV-2 gD fragment is as set forth in SEQ ID NO: 23. [0151] In some embodiments, an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-331 of gD (e.g., from HSV- 2 strain 333 or US6), as set forth in the following amino acid sequence: KYALADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVY YAVLERACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVME YTECPYNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKIN DWTEITQFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQR TVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVS SQIPPNWHIPSIQDVAPHH (SEQ ID NO: 5). [0152] In some embodiments, an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 5. In some embodiments, an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 5. [0153] In some embodiments, an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 30-331 of gD from HSV-2 (e.g., strain 333 or US6), as set forth in the following amino acid sequence: ADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVL ERACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECP 44 11979815v1 P-627574-PC YNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEI TQFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALY SLKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPN WHIPSIQDVAPHH (SEQ ID NO: 234). [0154] In some embodiments, an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 234. In some embodiments, an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 234. [0155] In some embodiments, an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 31-331 of gD from HSV-2 (e.g., strain 333 or US6), as set forth in the following amino acid sequence: DPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLE RACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECPY NKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEIT QFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALYS LKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPN WHIPSIQDVAPHH (SEQ ID NO: 110). [0156] In some embodiments, an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 110. In some embodiments, an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 110. [0157] In some embodiments, an HSV-2 gD fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-333 of gD from HSV-2 (e.g., strain 333 or US6), as set forth in the following amino acid sequence: DPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLE RACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECPY NKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEIT QFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALYS LKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPN WHIPSIQDVAPHHAP (SEQ ID NO: 120). 45 11979815v1 P-627574-PC [0158] In some embodiments, an HSV-2 gD fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 120. In some embodiments, an HSV-2 gD fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 120. [0159] In some embodiments, the full length HSV-2 gD encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: [0160] MGRLTSGVGTAALLVVAVGLRVVCAKYALADPSLKMADPNRFRGKNLPVLDQ LTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLERACRSVLLHAPSEAPQIVRGASDEA RKHTYNLTIAWYRMGDNCAIPITVMEYTECPYNKSLGVCPIRTQPRWSYYDSFSAVSED NLGFLMHAPAFETAGTYLRLVKINDWTEITQFILEHRARASCKYALPLRIPPAACLTSKA YQQGVTVDSIGMLPRFIPENQRTVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQPE LVPEDPEDSALLEDPAGTVSSQIPPNWHIPSIQDVAPHHAPAAPSNPGLIIGALAGSTLAV LVIGGIAFWVRRRAQMAPKRLRLPHIRDDDAPPSHQPLFY (SEQ ID NO: 6). [0161] In some embodiments, an HSV-2 gC comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 6. In some embodiments, an HSV-2 gC has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 6. [0162] In another embodiment, an HSV-2 gD, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in GenBank Accession Numbers: 1003204A, AAA45841.1, AAA45842.1, AAB60552.1, AAB60553.1, AAB60554.1, AAB60555.1, AAB72102.1, AAS01730.1, AAW23130.1, AAW23131.1, AAW23132.1, AAW23133.1, AAW23134.1, ABS84899.1, ABU45433.1, ABU45434.1, ABU45435.1, ABU45461.1, ABU45462.1, ACA28831.1, AEV91405.1, AFM93876.1, AFS18198.1, AFS18199.1, AFS18200.1, AFS18201.1, AFS18202.1, AFS18203.1, AFS18204.1, AFS18205.1, AFS18206.1, AFS18207.1, AFS18208.1, AFS18209.1, AFS18210.1, AFS18211.1, AFS18212.1, AFS18213.1, AFS18214.1, AFS18215.1, AFS18216.1, AFS18217.1, AFS18218.1, AFS18219.1, AFS18220.1, AFS18221.1, AHG54730.1, AIL27720.1, AIL27721.1, AIL27722.1, AIL27723.1, AIL27724.1, AIL27725.1, AIL27726.1, AIL27727.1, AIL27728.1, AIL27729.1, AIL27730.1, AIL27731.1, AIL28069.1, AIL28070.1, AKC42828.1, AKC59305.1, AKC59376.1, AKC59447.1, AKC59518.1, AKC59589.1, 46 11979815v1 P-627574-PC AMB66102.1, AMB66171.1, AMB66244.1, AMB66321.1, AMB66394.1, AMB66463.1, AQZ55754.1, AQZ55825.1, AQZ55896.1, AQZ55967.1, AQZ56038.1, AQZ56109.1, AQZ56180.1, AQZ56251.1, AQZ56322.1, AQZ56393.1, AQZ56464.1, AQZ56535.1, AQZ56606.1, AQZ56677.1, AQZ56748.1, AQZ56819.1, AQZ56890.1, AQZ56961.1, AQZ57032.1, AQZ57103.1, AQZ57174.1, AQZ57245.1, AQZ57316.1, AQZ57387.1, AQZ57458.1, AQZ57529.1, AQZ57600.1, AQZ57671.1, AQZ57742.1, AQZ57813.1, AQZ57884.1, AQZ57955.1, AQZ58026.1, AQZ58097.1, AQZ58168.1, AQZ58239.1, AQZ58310.1, AQZ58381.1, AQZ58452.1, AQZ58523.1, AQZ58594.1, AQZ58665.1, AQZ58736.1, AQZ58807.1, AQZ58878.1, AQZ58949.1, AQZ59020.1, AQZ59091.1, AQZ59162.1, ARO38000.1, ARO38001.1, ARO38002.1, ARO38003.1, ARO38004.1, ARO38005.1, ARO38006.1, ARO38007.1, ARO38008.1, ARO38009.1, ARO38010.1, ARO38011.1, ARO38012.1, ARO38013.1, ARO38014.1, ARO38015.1, ARO38016.1, ARO38017.1, ARO38018.1, ARO38019.1, ARO38020.1, ARO38021.1, ARO38022.1, ARO38023.1, ARO38024.1, ARO38025.1, ARO38026.1, ARO38027.1, ARO38028.1, ARO38029.1, ARO38030.1, ARO38031.1, ARO38032.1, ARO38033.1, ARO38034.1, ARO38035.1, ARO38036.1, ARO38037.1, ARO38038.1, ARO38039.1, ARO38040.1, ARO38041.1, ARO38042.1, ARO38043.1, ARO38044.1, CAA26025.1, CAB06713.1, CAC33573.1, CAT05432.1, P03172.2, Q69467.1, or YP_009137218.1. [0163] In another embodiment, the gD protein or fragment (e.g., immunogenic fragment) includes Y63. In another embodiment, the gD protein or fragment (e.g., immunogenic fragment) includes R159. In another embodiment, the gD protein or fragment (e.g., immunogenic fragment) includes D240. In another embodiment, the gD protein or fragment (e.g., immunogenic fragment) includes P246. In another embodiment, the gD protein or fragment (e.g., immunogenic fragment) includes a residue selected from Y63, R159, D240, and P246. In another embodiment, inclusion of one of these residues elicits antibodies that inhibit binding to nectin-1. [0164] The nomenclature used herein for gD amino acid residues includes the residues of the signal peptide encoded by the signal sequence. Thus, residue one of the mature protein is referred to as “26.” [0165] Each RNA encoding HSV-1 gD and HSV-2 gD protein or immunogenic fragment thereof represents a separate embodiment of the present disclosure. 47 11979815v1 P-627574-PC [0166] In some embodiments, an HSV gD, gC, and gE proteins, or immunogenic fragments thereof, encoded by the modified RNA as disclosed herein are described in US Patent Publication No.2013-0028925-A1, which is incorporated by reference herein in its entirety. [0167] In another embodiment, a gD protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure is an immunogenic fragment. In another embodiment, a gD immunoprotective antigen need not be the entire protein. The protective immune response generally involves, in another embodiment, an antibody response. In another embodiment, mutants, sequence conservative variants, and functional conservative variants of gD are useful in methods and compositions of the present disclosure, provided that all such variants retain the required immuno-protective effect. In another embodiment, the immunogenic fragment can comprise an immuno-protective gD antigen from any strain of HSV. In another embodiment, the immunogenic fragment can comprise sequence variants of HSV, as found in infected individuals. Glycoprotein C HSV-1 gC [0168] In another embodiment, a composition of the present disclosure comprises an RNA encoding HSV-1 gC protein. In another embodiment, a composition comprises an RNA encoding a fragment of an HSV-1 gC protein (e.g., an immunogenic fragment). [0169] In some embodiments, a nucleotide sequence of the RNA encoding an HSV-1 gC fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGGCCAUCUCCGG CGUGCCCGUGCUGGGCUUCUUCAUCAUCGCCGUGCUGAUGUCCGCCCAGG AGUCCUGGGCCGAGACCGCCUCCACCGGCCCCACCAUCACCGCCGGCGCCGUG ACCAACGCCUCCGAGGCCCCCACCUCCGGCUCCCCCGGCUCCGCCGCCUCCCCC GAGGUGACCCCCACCUCCACCCCCAACCCCAACAACGUGACCCAGAACAAGAC CACCCCCACCGAGCCCGCCUCCCCCCCCACCACCCCCAAGCCCACCUCCACCCC CAAGUCCCCCCCCACCUCCACCCCCGACCCCAAGCCCAAGAACAACACCACCCC CGCCAAGUCCGGCCGCCCCACCAAGCCCCCCGGCCCCGUGUGGUGCGACCGCCG CGACCCCCUGGCCCGCUACGGCUCCCGCGUGCAGAUCCGCUGCCGCUUCCGCA ACUCCACCCGCAUGGAGUUCCGCCUGCAGAUCUGGCGCUACUCCAUGGGCCCC UCCCCCCCCAUCGCCCCCGCCCCCGACCUGGAGGAGGUGCUGACCAACAUCACC 48 11979815v1 P-627574-PC GCCCCCCCCGGCGGCCUGCUGGUGUACGACUCCGCCCCCAACCUGACCGACCCC CACGUGCUGUGGGCCGAGGGCGCCGGCCCCGGCGCCGACCCCCCCCUGUACUC CGUGACCGGCCCCCUGCCCACCCAGCGCCUGAUCAUCGGCGAGGUGACCCCCG CCACCCAGGGCAUGUACUACCUGGCCUGGGGCCGCAUGGACUCCCCCCACGAG UACGGCACCUGGGUGCGCGUGCGCAUGUUCCGCCCCCCCUCCCUGACCCUGCA GCCCCACGCCGUGAUGGAGGGCCAGCCCUUCAAGGCCACCUGCACCGCCGCCG CCUACUACCCCCGCAACCCCGUGGAGUUCGACUGGUUCGAGGACGACCGCCAG GUGUUCAACCCCGGCCAGAUCGACACCCAGACCCACGAGCACCCCGACGGCUU CACCACCGUGUCCACCGUGACCUCCGAGGCCGUGGGCGGCCAGGUGCCCCCCC GCACCUUCACCUGCCAGAUGACCUGGCACCGCGACUCCGUGACCUUCUCCCGC CGCAACGCCACCGGCCUGGCCCUGGUGCUGCCCCGCCCCACCAUCACCAUGGA GUUCGGCGUGCGCCACGUGGUGUGCACCGCCGGCUGCGUGCCCGAGGGCGUGA CCUUCGCCUGGUUCCUGGGCGACGACCCCUCCCCCGCCGCCAAGUCCGCCGUG ACCGCCCAGGAGUCCUGCGACCACCCCGGCCUGGCCACCGUGCGCUCCACCCU GCCCAUCUCCUACGACUACUCCGAGUACAUCUGCCGCCUGACCGGCUACCCCG CCGGCAUCCCCGUGCUGGAGCACCACUAACUAGUAGUGACUGACUAGGAUCUGG UUACCACUAAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUAC CAACUUACACUUACAAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCC UAAUAAAAAGAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAC (SEQ ID NO: 7) [0170] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 21) to assist expression of the HSV-1 gC fragment. In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264). [0171] In another embodiment, a nucleotide sequence of the RNA encoding an HSV-1 gC fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or any combinations thereof. In some embodiments, the sequence of the HSV-1 gC fragment is as set forth in SEQ ID NO: 24. 49 11979815v1 P-627574-PC [0172] In some embodiments, an HSV-1 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 27-457 of gC from HSV-1 (e.g., KOS strain), as set forth in the following amino acid sequence: ETASTGPTITAGAVTNASEAPTSGSPGSAASPEVTPTSTPNPNNVTQNKTTPTEPASPP TTPKPTSTPKSPPTSTPDPKPKNNTTPAKSGRPTKPPGPVWCDRRDPLARYGSRVQIR CRFRNSTRMEFRLQIWRYSMGPSPPIAPAPDLEEVLTNITAPPGGLLVYDSAPNLTDP HVLWAEGAGPGADPPLYSVTGPLPTQRLIIGEVTPATQGMYYLAWGRMDSPHEYGT WVRVRMFRPPSLTLQPHAVMEGQPFKATCTAAAYYPRNPVEFDWFEDDRQVFNPG QIDTQTHEHPDGFTTVSTVTSEAVGGQVPPRTFTCQMTWHRDSVTFSRRNATGLAL VLPRPTITMEFGVRHVVCTAGCVPEGVTFAWFLGDDPSPAAKSAVTAQESCDHPGL ATVRSTLPISYDYSEYICRLTGYPAGIPVLEHH (SEQ ID NO: 8). [0173] In some embodiments, an HSV-1 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 8. In some embodiments, an HSV-1 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 8. [0174] In some embodiments, the HSV-1 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 25-457 of gC from HSV-1 (e.g., KOS strain), as set forth in the following amino acid sequence: GSETASTGPTITAGAVTNASEAPTSGSPGSAASPEVTPTSTPNPNNVTQNKTTPTEPAS PPTTPKPTSTPKSPPTSTPDPKPKNNTTPAKSGRPTKPPGPVWCDRRDPLARYGSRVQ IRCRFRNSTRMEFRLQIWRYSMGPSPPIAPAPDLEEVLTNITAPPGGLLVYDSAPNLTD PHVLWAEGAGPGADPPLYSVTGPLPTQRLIIGEVTPATQGMYYLAWGRMDSPHEYG TWVRVRMFRPPSLTLQPHAVMEGQPFKATCTAAAYYPRNPVEFDWFEDDRQVFNP GQIDTQTHEHPDGFTTVSTVTSEAVGGQVPPRTFTCQMTWHRDSVTFSRRNATGLA LVLPRPTITMEFGVRHVVCTAGCVPEGVTFAWFLGDDPSPAAKSAVTAQESCDHPG LATVRSTLPISYDYSEYICRLTGYPAGIPVLEHH (SEQ ID NO: 240). [0175] In some embodiments, an HSV-1 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 240. In some embodiments, an HSV-1 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 240. 50 11979815v1 P-627574-PC [0176] In some embodiments, a gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 27-457 of gC from an HSV-1 strain. [0177] In some embodiments, the full length HSV-1 gC encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MAPGRVGLAVVLWGLLWLGAGVAGGSETASTGPTITAGAVTNASEAPTSGSPGSA ASPEVTPTSTPNPNNVTQNKTTPTEPASPPTTPKPTSTPKSPPTSTPDPKPKNNTTPAKS GRPTKPPGPVWCDRRDPLARYGSRVQIRCRFRNSTRMEFRLQIWRYSMGPSPPIAPA PDLEEVLTNITAPPGGLLVYDSAPNLTDPHVLWAEGAGPGADPPLYSVTGPLPTQRLI IGEVTPATQGMYYLAWGRMDSPHEYGTWVRVRMFRPPSLTLQPHAVMEGQPFKAT CTAAAYYPRNPVEFDWFEDDRQVFNPGQIDTQTHEHPDGFTTVSTVTSEAVGGQVP PRTFTCQMTWHRDSVTFSRRNATGLALVLPRPTITMEFGVRHVVCTAGCVPEGVTF AWFLGDDPSPAAKSAVTAQESCDHPGLATVRSTLPISYDYSEYICRLTGYPAGIPVLE HHGSHQPPPRDPTERQVIEAIEWVGIGIGVLAAGVLVVTAIVYVVRTSQSRQRHRR (SEQ ID NO:9). [0178] In some embodiments, an HSV-2 gC comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 9. In some embodiments, an HSV-2 gC has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 9. [0179] In another embodiment, an HSV-1 gC encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: AAA45779.1, AAA96680.1, ABI63505.1, ABM52973.1, ABM52976.1, ABM52977.1, ACM62267.1, ADD60042.1, ADD60119.1, ADM22367.1, ADM22444.1, ADM22520.1, ADM22597.1, ADM22674.1, ADM22751.1, ADM22827.1, ADM22904.1, ADM22981.1, ADM23057.1, ADM23133.1, ADM23210.1, ADM23287.1, ADM23361.1, ADM23435.1, ADM23509.1, ADM23583.1, ADM23658.1, ADM23733.1, ADM23809.1, AEQ77075.1, AEQ77099.1, AER37628.1, AER37697.1, AER37767.1, AER37838.1, AER37910.1, AER37981.1, AER38051.2, AFA36179.1, AFA36180.1, AFA36181.1, AFA36182.1, AFA36183.1, AFA36184.1, AFA36185.1, AFA36186.1, AFA36187.1, AFA36188.1, AFA36189.1, AFA36190.1, AFA36191.1, AFA36192.1, AFA36193.1, AFA36194.1, AFA36195.1, AFA36196.1, AFA36197.1, AFA36198.1, AFA36199.1, AFA36200.1, AFA36201.1, AFA36202.1, AFA36203.1, 51 11979815v1 P-627574-PC AFE62872.1, AFH78104.1, AFI23635.1, AFK50391.1, AFP86408.1, AGZ01906.1, AIR95840.1, AJE59989.1, AJE60060.1, AJE60131.1, AJE60202.1, AKE48623.1, AKE98415.1, AKE98416.1, AKE98417.1, AKE98418.1, AKE98419.1, AKE98420.1, AKE98421.1, AKE98422.1, AKE98423.1, AKE98424.1, AKE98425.1, AKE98426.1, AKE98427.1, AKE98428.1, AKE98429.1, AKE98430.1, AKE98431.1, AKE98432.1, AKE98433.1, AKE98434.1, AKE98435.1, AKG59227.1, AKG59299.1, AKG59372.1, AKG59444.1, AKG59516.1, AKG59591.1, AKG59663.1, AKG59736.1, AKG59807.1, AKG59879.1, AKG59953.1, AKG60027.1, AKG60099.1, AKG60170.1, AKG60243.1, AKG60316.1, AKG60386.1, AKG60456.1, AKG60528.1, AKG60601.1, AKG60674.1, AKG60745.1, AKG60817.1, AKG60887.1, AKG60959.1, AKG61032.1, AKG61104.1, AKG61175.1, AKG61248.1, AKG61321.1, AKG61392.1, AKG61464.1, AKG61537.1, AKG61611.1, AKG61684.1, AKG61756.1, AKG61828.1, AKG61902.1, AKG61974.1, AKH80444.1, AKH80517.1, AKM76368.1, ALM22613.1, ALM22687.1, ALM22761.1, ALM22835.1, ALO18641.1, ALO18717.1, AMB65642.1, AMB65715.1, AMB65862.1, AMN09813.1, ANN83942.1, ANN84019.1, ANN84095.1, ANN84172.1, ANN84249.1, ANN84326.1, ANN84403.1, ANN84478.1, ANN84555.1, ANN84632.1, ANN84708.1, ANN84785.1, ANN84861.1, ANN84938.1, ANN85014.1, ANN85091.1, ANN85167.1, ANN85242.1, ANN85319.1, ANN85396.1, ANN85472.1, ANN85549.1, ANN85626.1, ANN85703.1, ANN85779.1, AOY34308.1, AOY36663.1, AOY36687.1, ARB08935.1, ARO38059.1, ARO38060.1, ARO38061.1, ARO38062.1, ARO38063.1, ARO38064.1, ARO38065.1, ARO38066.1, ASM47642.1, ASM47719.1, ASM47796.1, ASM47871.1, BAM73394.1, CAA32294.1, CAB40083.1, CAD13356.1, CAD13357.1, CAD13358.1, CAD13359.1, CAD13360.1, CAD13361.1, CAD13362.1, CAD13363.1, CAD13364.1, CAD13365.1, CAD13366.1, CAD13367.1, CAD13368.1, CAD13369.1, CAD13370.1, CAD13371.1, CAD13372.1, CAD13373.1, CAD13374.1, CAD13375.1, CAD13376.1, CAD13377.1, CAD13378.1, P04290.1, P04488.1, P09855.1, P10228.1, P28986.1, SBO07729.1, SBO07793.1, SBO07798.1, SBO07812.1, SBO07880.1, SBS69375.1, SBS69379.1, SBS69440.1, SBS69448.1, SBS69560.1, SBS69599.1, SBS69602.1, SBS69637.1, SBS69790.1, SBT69374.1, SCL76887.1, YP_009137119.1, or YP_009137143.1. 52 11979815v1 P-627574-PC HSV-2 gC [0180] In some embodiments, a composition comprises an RNA encoding an HSV-2 gC protein. In another embodiment, a composition comprises an RNA encoding a fragment of an HSV-2 gC protein (e.g., immunogenic fragment). [0181] In some embodiments, a nucleotide sequence of the RNA encoding an HSV-2 gC fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCU GCUGCUGCUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCGCCUCCCC CGGCCGCACCAUCACCGUGGGCCCCCGCGGCAACGCCUCCAACGCCGCCCCCUC CGCCUCCCCCCGCAACGCCUCCGCCCCCCGCACCACCCCCACCCCCCCCCAGCCC CGCAAGGCCACCAAGUCCAAGGCCUCCACCGCCAAGCCCGCCCCCCCCCCCAAG ACCGGCCCCCCCAAGACCUCCUCCGAGCCCGUGCGCUGCAACCGCCACGACCCC CUGGCCCGCUACGGCUCCCGCGUGCAGAUCCGCUGCCGCUUCCCCAACUCCAC CCGCACCGAGUUCCGCCUGCAGAUCUGGCGCUACGCCACCGCCACCGACGCCG AGAUCGGCACCGCCCCCUCCCUGGAGGAGGUGAUGGUGAACGUGUCCGCCCCC CCCGGCGGCCAGCUGGUGUACGACUCCGCCCCCAACCGCACCGACCCCCACGU GAUCUGGGCCGAGGGCGCCGGCCCCGGCGCCUCCCCCCGCCUGUACUCCGUGG UGGGCCCCCUGGGCCGCCAGCGCCUGAUCAUCGAGGAGCUGACCCUGGAGACC CAGGGCAUGUACUACUGGGUGUGGGGCCGCACCGACCGCCCCUCCGCCUACGG CACCUGGGUGCGCGUGCGCGUGUUCCGCCCCCCCUCCCUGACCAUCCACCCCCA CGCCGUGCUGGAGGGCCAGCCCUUCAAGGCCACCUGCACCGCCGCCACCUACU ACCCCGGCAACCGCGCCGAGUUCGUGUGGUUCGAGGACGGCCGCCGCGUGUUC GACCCCGCCCAGAUCCACACCCAGACCCAGGAGAACCCCGACGGCUUCUCCAC CGUGUCCACCGUGACCUCCGCCGCCGUGGGCGGCCAGGGCCCCCCCCGCACCU UCACCUGCCAGCUGACCUGGCACCGCGACUCCGUGUCCUUCUCCCGCCGCAAC GCCUCCGGCACCGCCUCCGUGCUGCCCCGCCCCACCAUCACCAUGGAGUUCACC GGCGACCACGCCGUGUGCACCGCCGGCUGCGUGCCCGAGGGCGUGACCUUCGC CUGGUUCCUGGGCGACGACUCCUCCCCCGCCGAGAAGGUGGCCGUGGCCUCCC AGACCUCCUGCGGCCGCCCCGGCACCGCCACCAUCCGCUCCACCCUGCCCGUGU CCUACGAGCAGACCGAGUACAUCUGCCGCCUGGCCGGCUACCCCGACGGCAUC 53 11979815v1 P-627574-PC CCCGUGCUGGAGCACCACUAACUAGUAGUGACUGACUAGGAUCUGGUUACCACU AAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUAC ACUUACAAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAAAA GAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA C (SEQ ID NO: 10). [0182] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 19) to assist expression of the HSV-2 gC fragment. In some embodiments, italicized residues represent 3’ untranslated sequences and a poly adenylation tail (SEQ ID NO: 244). [0183] In another embodiment, a nucleotide sequence of the RNA encoding an HSV-2 gC fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or any combinations thereof. In some embodiments, the sequence of the HSV-2 gC fragment is as set forth in SEQ ID NO: 25. [0184] In some embodiments, an HSV-2 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 27-426 of gC from HSV-2 (e.g. strain 333 or UL44), as set forth in the following amino acid sequence: ASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKT GPPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTESRLQIWRYATATDAEIGTAP SLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRL IIEELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATC TAATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGPP RTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFA WFLGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEH H (SEQ ID NO: 11). [0185] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 11. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 11. 54 11979815v1 P-627574-PC [0186] In some embodiments, an HSV-2 gC fragment comprises the following amino acid sequence: ASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKT GPPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTEFRLQIWRYATATDAEIGTAP SLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRL IIEELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATC TAATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGPP RTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFA WFLGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEH H (SEQ ID NO: 28). [0187] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 28. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 28. [0188] In some embodiments, the HSV-2 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 28-426 of gC from HSV-2 (e.g., strain 333 or UL44), as set forth in the following amino acid sequence: SPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTG PPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTESRLQIWRYATATDAEIGTAPS LEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLII EELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCT AATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGPPR TFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFAWF LGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEHH (SEQ ID NO: 223). [0189] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 223. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 223. 55 11979815v1 P-627574-PC [0190] In some embodiments, an HSV-2 gC fragment comprises the following amino acid sequence: SPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPKTG PPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTEFRLQIWRYATATDAEIGTAPS LEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQRLII EELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKATCT AATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGPPR TFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFAWF LGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEHH (SEQ ID NO: 224). [0191] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 224. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 224. [0192] In some embodiments, the HSV-2 gC fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 26-426 of gC from HSV-2 (e.g., strain 333 or UL44), as set forth in the following amino acid sequence: SASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPK TGPPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTESRLQIWRYATATDAEIGTA PSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQR LIIEELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKAT CTAATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGP PRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFA WFLGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEH H (SEQ ID NO: 241). [0193] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 241. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 241. 56 11979815v1 P-627574-PC [0194] In some embodiments, an HSV-2 gC fragment comprises the following amino acid sequence: SASPGRTITVGPRGNASNAAPSASPRNASAPRTTPTPPQPRKATKSKASTAKPAPPPK TGPPKTSSEPVRCNRHDPLARYGSRVQIRCRFPNSTRTEFRLQIWRYATATDAEIGTA PSLEEVMVNVSAPPGGQLVYDSAPNRTDPHVIWAEGAGPGASPRLYSVVGPLGRQR LIIEELTLETQGMYYWVWGRTDRPSAYGTWVRVRVFRPPSLTIHPHAVLEGQPFKAT CTAATYYPGNRAEFVWFEDGRRVFDPAQIHTQTQENPDGFSTVSTVTSAAVGGQGP PRTFTCQLTWHRDSVSFSRRNASGTASVLPRPTITMEFTGDHAVCTAGCVPEGVTFA WFLGDDSSPAEKVAVASQTSCGRPGTATIRSTLPVSYEQTEYICRLAGYPDGIPVLEH H (SEQ ID NO: 242). [0195] In some embodiments, an HSV-2 gC fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 242. In some embodiments, an HSV-2 gC fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 242. [0196] In some embodiments, the full length HSV-2 gC encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MALGRVGLAVGLWGLLWVGVVVVLANASPGRTITVGPRGNASNAAPSASPRNASA PRTTPTPPQPRKATKSKASTAKPAPPPKTGPPKTSSEPVRCNRHDPLARYGSRVQIRC RFPNSTRTEFRLQIWRYATATDAEIGTAPSLEEVMVNVSAPPGGQLVYDSAPNRTDP HVIWAEGAGPGASPRLYSVVGPLGRQRLIIEELTLETQGMYYWVWGRTDRPSAYGT WVRVRVFRPPSLTIHPHAVLEGQPFKATCTAATYYPGNRAEFVWFEDGRRVFDPAQI HTQTQENPDGFSTVSTVTSAAVGGQGPPRTFTCQLTWHRDSVSFSRRNASGTASVLP RPTITMEFTGDHAVCTAGCVPEGVTFAWFLGDDSSPAEKVAVASQTSCGRPGTATIR STLPVSYEQTEYICRLAGYPDGIPVLEHHGSHQPPPRDPTERQVIRAVEGAGIGVAVL VAVVLAGTAVVYLTHASSVRYRRLR (SEQ ID NO: 12). [0197] In some embodiments, an HSV-2 gC comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 12. In some embodiments, an HSV-2 gC has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 12. 57 11979815v1 P-627574-PC [0198] In some embodiments, an HSV-2 gC encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: AAA20532.1, AAA66442.1, AAB60549.1, AAB60550.1, AAB60551.1, AAB72101.1, ABU45429.1, ABU45430.1, ABU45431.1, ABU45432.1, ABU45459.1, ABU45460.1, AEV91348.1, AEV91383.1, AEV91407.1, AFM93864.1, AHG54708.1, AKC42808.1, AKC59285.1, AKC59357.1, AKC59428.1, AKC59499.1, AKC59570.1, AMB66008.1, AMB66079.1, AMB66151.1, AMB66224.1, AMB66252.1, AMB66253.1, AMB66368.1, AMB66441.1, AQZ55735.2, AQZ55806.1, AQZ55877.1, AQZ55948.1, AQZ56019.1, AQZ56090.1, AQZ56161.2, AQZ56232.2, AQZ56303.2, AQZ56374.2, AQZ56445.1, AQZ56516.1, AQZ56587.1, AQZ56658.1, AQZ56729.2, AQZ56800.1, AQZ56871.1, AQZ56942.2, AQZ57013.1, AQZ57084.2, AQZ57155.1, AQZ57226.1, AQZ57297.1, AQZ57368.1, AQZ57439.1, AQZ57510.1, AQZ57581.1, AQZ57652.1, AQZ57723.1, AQZ57794.2, AQZ57865.2, AQZ57936.1, AQZ58007.2, AQZ58078.1, AQZ58149.2, AQZ58220.1, AQZ58291.1, AQZ58362.1, AQZ58433.1, AQZ58504.1, AQZ58575.1, AQZ58646.1, AQZ58717.2, AQZ58788.2, AQZ58859.2, AQZ58930.1, AQZ59001.2, AQZ59072.1, AQZ59143.1, ARO38067.1, ARO38068.1, ARO38069.1, ARO38070.1, ARO38071.1, ARO38072.1, CAA25687.1, CAA26025.1, CAB06730.1, CAB06734.1, CAB96544.1, P03173.1, P06475.1, P89475.1, Q89730.1, YP_009137161.1, YP_009137196.1, or YP_009137220.1. [0199] In other embodiments, an HSV gC protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises a properdin interfering domain “Properdin-interfering domain” refers, in some embodiments, to a domain that blocks or inhibits binding of a host C3b molecule with a host properdin molecule. In another embodiment, the term refers to a domain that blocks or inhibits an interaction of a host C3b molecule with a host properdin molecule. [0200] In another embodiment, an HSV gC protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure is a C5 interfering domain. In another embodiment, the gC protein fragment is a portion of a C5 interfering domain. “C5-interfering domain” refers, in another embodiment, to a domain that interferes with binding of a host C3b molecule with a host C5 molecule. In another embodiment, the term refers to a domain that interferes with the interaction of a host C3b molecule with a host C5 molecule. 58 11979815v1 P-627574-PC [0201] Each RNA encoding HSV-1 gCor HSV-2 gC protein or fragment thereof represents a separate embodiment of the present disclosure. [0202] In another embodiment, a gC protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure is an immunogenic fragment. In another embodiment, a gC immunoprotective antigen need not be the entire protein. The protective immune response generally involves, in another embodiment, an antibody response. In another embodiment, mutants, sequence conservative variants, and functional conservative variants of gC are useful in methods and compositions of the present disclosure, provided that all such variants retain the required immuno-protective effect. In another embodiment, the immunogenic fragment can comprise an immuno-protective gC antigen from any strain of HSV. In another embodiment, the immunogenic fragment can comprise sequence variants of HSV, as found in infected individuals. Glycoprotein E HSV-1 gE [0203] In another embodiment, a composition of the present disclosure comprises an RNA encoding HSV-1 gE protein. In another embodiment, a composition comprises an RNA encoding a fragment of an HSV-1 gE protein (e.g., immunogenic fragment). [0204] In some embodiments, a nucleotide sequence of the RNA encoding an HSV-1 gE fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCU GCUGCUGCUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCAAGACCUC CUGGCGCCGCGUGUCCGUGGGCGAGGACGUGUCCCUGCUGCCCGCCCCCGGCC CCACCGGCCGCGGCCCCACCCAGAAGCUGCUGUGGGCCGUGGAGCCCCUGGAC GGCUGCGGCCCCCUGCACCCCUCCUGGGUGUCCCUGAUGCCCCCCAAGCAGGU GCCCGAGACCGUGGUGGACGCCGCCUGCAUGCGCGCCCCCGUGCCCCUGGCCA UGGCCUACGCCCCCCCCGCCCCCUCCGCCACCGGCGGCCUGCGCACCGACUUCG UGUGGCAGGAGCGCGCCGCCGUGGUGAACCGCUCCCUGGUGAUCUACGGCGUG CGCGAGACCGACUCCGGCCUGUACACCCUGUCCGUGGGCGACAUCAAGGACCC CGCCCGCCAGGUGGCCUCCGUGGUGCUGGUGGUGCAGCCCGCCCCCGUGCCCA CCCCCCCCCCCACCCCCGCCGACUACGACGAGGACGACAACGACGAGGGCGAG GGCGAGGACGAGUCCCUGGCCGGCACCCCCGCCUCCGGCACCCCCCGCCUGCCC 59 11979815v1 P-627574-PC CCCUCCCCCGCCCCCCCCCGCUCCUGGCCCUCCGCCCCCGAGGUGUCCCACGUG CGCGGCGUGACCGUGCGCAUGGAGACCCCCGAGGCCAUCCUGUUCUCCCCCGG CGAGGCCUUCUCCACCAACGUGUCCAUCCACGCCAUCGCCCACGACGACCAGA CCUACACCAUGGACGUGGUGUGGCUGCGCUUCGACGUGCCCACCUCCUGCGCC GAGAUGCGCAUCUACGAGUCCUGCCUGUACCACCCCCAGCUGCCCGAGUGCCU GUCCCCCGCCGACGCCCCCUGCGCCGCCUCCACCUGGACCUCCCGCCUGGCCGU GCGCUCCUACGCCGGCUGCUCCCGCACCAACCCCCCCCCCCGCUGCUCCGCCGA GGCCCACAUGGAGCCCUUCCCCGGCCUGGCCUGGCAGGCCGCCUCCGUGAACC UGGAGUUCCGCGACGCCUCCCCCCAGCACUCCGGCCUGUACCUGUGCGUGGUG UACGUGAACGACCACAUCCACGCCUGGGGCCACAUCACCAUCAACACCGCCGC CCAGUACCGCAACGCCGUGGUGGAGCAGCCCCUGCCCCAGCGCGGCGCCGACC UGGCCGAGCCCACCCACCCCCACGUGGGCGCCUAACUAGUAGUGACUGACUAGG AUCUGGUUACCACUAAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACA UAAUACCAACUUACACUUACAAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUC UGCUCCUAAUAAAAAGAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAC (SEQ ID NO: 13). [0205] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 19) to assist expression of the HSV-1 gE fragment. In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264). [0206] In another embodiment, a nucleotide sequence of the RNA encoding an HSV-1 gE fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof. In some embodiments, the sequence of the HSV-1 gE fragment is as set forth in SEQ ID NO: 26. [0207] In some embodiments, an HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-409 of gE from HSV-1 (e.g., NS strain), as set forth in the following amino acid sequence: KTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLWAVEPLDGCGPLHPSWVSLMPPKQV PETVVDAACMRAPVPLAMAYAPPAPSATGGLRTDFVWQERAAVVNRSLVIYGVRE TDSGLYTLSVGDIKDPARQVASVVLVVQPAPVPTPPPTPADYDEDDNDEGEGEDESL 60 11979815v1 P-627574-PC AGTPASGTPRLPPSPAPPRSWPSAPEVSHVRGVTVRMETPEAILFSPGEAFSTNVSIHA IAHDDQTYTMDVVWLRFDVPTSCAEMRIYESCLYHPQLPECLSPADAPCAASTWTS RLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLAWQAASVNLEFRDASPQHSGLYLCV VYVNDHIHAWGHITINTAAQYRNAVVEQPLPQRGADLAEPTHPHVGA (SEQ ID NO: 14). [0208] In some embodiments, an HSV-1 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 14. In some embodiments, an HSV-1 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 14. [0209] In some embodiments, a gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-409 of gE from an HSV-1 strain (SEQ ID NO: 14). [0210] In some embodiments, an HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 21-409 of gE from HSV-1 (e.g., NS strain or US8), as set forth in the following amino acid sequence: GTPKTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLWAVEPLDGCGPLHPSWVSLMPP KQVPETVVDAACMRAPVPLAMAYAPPAPSATGGLRTDFVWQERAAVVNRSLVIYG VRETDSGLYTLSVGDIKDPARQVASVVLVVQPAPVPTPPPTPADYDEDDNDEGEGE DESLAGTPASGTPRLPPSPAPPRSWPSAPEVSHVRGVTVRMETPEAILFSPGEAFSTNV SIHAIAHDDQTYTMDVVWLRFDVPTSCAEMRIYESCLYHPQLPECLSPADAPCAAST WTSRLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLAWQAASVNLEFRDASPQHSGLY LCVVYVNDHIHAWGHITINTAAQYRNAVVEQPLPQRGADLAEPTHPHVGA (SEQ ID NO: 248). [0211] In some embodiments, an HSV-1 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 248. In some embodiments, an HSV-1 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 248. [0212] In some embodiments, an HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 21-409 of gE from an HSV-1 strain. 61 11979815v1 P-627574-PC [0213] In some embodiments, an HSV-1 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-309 of gE from HSV-1 (e.g., NS strain or US8), as set forth in the following amino acid sequence: GTPKTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLWAVEPLDGCGPLHPSWVSLMPP KQVPETVVDAACMRAPVPLAMAYAPPAPSATGGLRTDFVWQERAAVVNRSLVIYG VRETDSGLYTLSVGDIKDPARQVASVVLVVQPAPVPTPPPTPADYDEDDNDEGEGE DESLAGTPASGTPRLPPSPAPPRSWPSAPEVSHVRGVTVRMETPEAILFSPGEAFSTNV SIHAIAHDDQTYTMDVVWLRFDVPTSCAEMRIYESCLYHPQLPECLSPADAPCAAST WTSRLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLAWQAASVNLEFRDASPQHSGLY LCVVYVNDHIHAWGHITINTAAQYRNAVVEQPLPQRGADLAEPTHPHVGA (SEQ ID NO: 248). [0214] In some embodiments, an HSV-1 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 248. In some embodiments, an HSV-1 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 248. [0215] In some embodiments, a full length HSV-1 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MDRGAVVGFLLGVCVVSCLAGTPKTSWRRVSVGEDVSLLPAPGPTGRGPTQKLLW AVEPLDGCGPLHPSWVSLMPPKQVPETVVDAACMRAPVPLAMAYAPPAPSATGGL RTDFVWQERAAVVNRSLVIYGVRETDSGLYTLSVGDIKDPARQVASVVLVVQPAPV PTPPPTPADYDEDDNDEGEGEDESLAGTPASGTPRLPPSPAPPRSWPSAPEVSHVRGV TVRMETPEAILFSPGEAFSTNVSIHAIAHDDQTYTMDVVWLRFDVPTSCAEMRIYES CLYHPQLPECLSPADAPCAASTWTSRLAVRSYAGCSRTNPPPRCSAEAHMEPFPGLA WQAASVNLEFRDASPQHSGLYLCVVYVNDHIHAWGHITINTAAQYRNAVVEQPLP QRGADLAEPTHPHVGAPPHAPPTHGALRLGAVMGAALLLSALGLSVWACMTCWR RRAWRAVKSRASGKGPTYIRVADSELYADWSSDSEGERDQVPWLAPPERPDSPSTN GSGFEILSPTAPSVYPRSDGHQSRRQLTTFGSGRPDRRYSQASDSSVFW (SEQ ID NO: 15). [0216] In some embodiments, an HSV-1 gE comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ 62 11979815v1 P-627574-PC ID NO: 15. In some embodiments, an HSV-1 gE has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO:15. [0217] In another embodiment, an HSV-1 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: AAA45779.1, AAA96680.1, ABI63526.1, ACM62297.1, ADD60055.1, ADD60132.1, ADM22391.1, ADM22468.1, ADM22544.1, ADM22621.1, ADM22698.1, ADM22775.1, ADM22851.1, ADM22928.1, ADM23005.1, ADM23081.1, ADM23157.1, ADM23233.1, ADM23311.1, ADM23385.1, ADM23459.1, ADM23533.1, ADM23607.1, ADM23682.1, ADM23757.1, ADM23833.1, ADN34689.1, ADN34692.1, ADN34695.1, AEQ77099.1, AER37649.1, AER37717.1, AER37788.1, AER37859.1, AER37931.1, AER38002.1, AER38072.1, AFA36179.1, AFA36180.1, AFA36181.1, AFA36182.1, AFA36183.1, AFA36184.1, AFA36185.1, AFA36186.1, AFA36187.1, AFA36188.1, AFA36189.1, AFA36190.1, AFA36191.1, AFA36192.1, AFA36193.1, AFA36194.1, AFA36195.1, AFA36196.1, AFA36197.1, AFA36198.1, AFA36199.1, AFA36200.1, AFA36201.1, AFA36202.1, AFA36203.1, AFE62896.1, AFI23659.1, AFK50417.1, AFP86432.1, AGZ01930.1, AIR95859.1, AJE60011.1, AJE60082.1, AJE60153.1, AJE60224.1, AJE60295.1, AKE48647.1, AKE98373.1, AKE98374.1, AKE98375.1, AKE98376.1, AKE98377.1, AKE98378.1, AKE98379.1, AKE98380.1, AKE98381.1, AKE98382.1, AKE98383.1, AKE98384.1, AKE98385.1, AKE98386.1, AKE98387.1, AKE98388.1, AKE98389.1, AKE98390.1, AKE98391.1, AKE98392.1, AKE98393.1, AKG59248.1, AKG59320.1, AKG59393.1, AKG59464.1, AKG59538.1, AKG59611.1, AKG59684.1, AKG59757.1, AKG59828.1, AKG59900.1, AKG59974.1, AKG60048.1, AKG60120.1, AKG60191.1, AKG60263.1, AKG60336.1, AKG60406.1, AKG60476.1, AKG60548.1, AKG60622.1, AKG60694.1, AKG60765.1, AKG60837.1, AKG60908.1, AKG60980.1, AKG61052.1, AKG61125.1, AKG61196.1, AKG61269.1, AKG61341.1, AKG61413.1, AKG61486.1, AKG61558.1, AKG61631.1, AKG61705.1, AKG61776.1, AKG61849.1, AKG61922.1, AKG61995.1, AKH80465.1, AKH80538.1, ALM22637.1, ALM22711.1, ALM22785.1, ALM22859.1, ALO18664.1, ALO18740.1, AMB65664.1, AMB65737.1, AMB65811.1, AMB65887.1, AMB65958.1, AMN09834.1, ANN83966.1, ANN84043.1, ANN84119.1, ANN84196.1, ANN84273.1, ANN84350.1, ANN84426.1, ANN84502.1, ANN84579.1, ANN84655.1, ANN84732.1, ANN84808.1, ANN84885.1, ANN84961.1, ANN85038.1, ANN85114.1, ANN85189.1, ANN85266.1, 63 11979815v1 P-627574-PC ANN85343.1, ANN85418.1, ANN85496.1, ANN85573.1, ANN85650.1, ANN85726.1, ANN85803.1, AOY34085.1, AOY36687.1, ARB08959.1, ARO38073.1, ARO38074.1, ARO38075.1, ARO38076.1, ARO38077.1, ARO38078.1, ARO38079.1, ARO38080.1, ASM47642.1, ASM47666.1, ASM47743.1, ASM47820.1, ASM47895.1, BAM73421.1, CAA26062.1, CAA32272.1, CAF24756.1, CAF24757.1, CAF24758.1, CAF24759.1, CAF24760.1, CAF24761.1, CAF24762.1, CAF24763.1, CAF24764.1, CAF24765.1, CAF24766.1, CAF24767.1, CAF24768.1, CAF24769.1, CAF24770.1, CAF24771.1, CAF24772.1, CAF24773.1, CAF24774.1, CAF24775.1, CAF24776.1, CAF24777.1, CAF24778.1, CAF24779.1, CAF24780.1, CAF24781.1, CAF24782.1, CAF24783.1, CAF24784.1, CAF24785.1, P04290.1, P04488.1, P28986.1, Q703F0.1, SBO07910.1, SBS69571.1, SBS69576.1, SBS69595.1, SBS69636.1, SBS69693.1, SBS69701.1, SBS69722.1, SBS69732.1, SBS69813.1, SBT69397.1, or YP_009137143.1. HSV-2 gE [0218] In some embodiments, a composition comprises an RNA encoding an HSV-2 gE protein. In another embodiment, a composition comprises an RNA encoding a fragment of an HSV-2 gE protein (e.g., immunogenic fragment). [0219] In some embodiments, a nucleotide sequence of the RNA encoding an HSV-2 gE fragment comprises: GGAAUAAAAGUCUCAACACAACAUAUACAAAACAAACGAAUCUCAAGCAAUC AAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAG CAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCAUGCGCAUGCAGCU GCUGCUGCUGAUCGCCCUGUCCCUGGCCCUGGUGACCAACUCCCGCACCUC CUGGAAGCGCGUGACCUCCGGCGAGGACGUGGUGCUGCUGCCCGCCCCCGCCG GCCCCGAGGAGCGCACCCGCGCCCACAAGCUGCUGUGGGCCGCCGAGCCCCUG GACGCCUGCGGCCCCCUGCGCCCCUCCUGGGUGGCCCUGUGGCCCCCCCGCCGC GUGCUGGAGACCGUGGUGGACGCCGCCUGCAUGCGCGCCCCCGAGCCCCUGGC CAUCGCCUACUCCCCCCCCUUCCCCGCCGGCGACGAGGGCCUGUACUCCGAGC UGGCCUGGCGCGACCGCGUGGCCGUGGUGAACGAGUCCCUGGUGAUCUACGGC GCCCUGGAGACCGACUCCGGCCUGUACACCCUGUCCGUGGUGGGCCUGUCCGA CGAGGCCCGCCAGGUGGCCUCCGUGGUGCUGGUGGUGGAGCCCGCCCCCGUGC CCACCCCCACCCCCGACGACUACGACGAGGAGGACGACGCCGGCGUGUCCGAG CGCACCCCCGUGUCCGUGCCCCCCCCCACCCCCCCCCGCCGCCCCCCCGUGGCC 64 11979815v1 P-627574-PC CCCCCCACCCACCCCCGCGUGAUCCCCGAGGUGUCCCACGUGCGCGGCGUGACC GUGCACAUGGAGACCCCCGAGGCCAUCCUGUUCGCCCCCGGCGAGACCUUCGG CACCAACGUGUCCAUCCACGCCAUCGCCCACGACGACGGCCCCUACGCCAUGG ACGUGGUGUGGAUGCGCUUCGACGUGCCCUCCUCCUGCGCCGAGAUGCGCAUC UACGAGGCCUGCCUGUACCACCCCCAGCUGCCCGAGUGCCUGUCCCCCGCCGA CGCCCCCUGCGCCGUGUCCUCCUGGGCCUACCGCCUGGCCGUGCGCUCCUACG CCGGCUGCUCCCGCACCACCCCCCCCCCCCGCUGCUUCGCCGAGGCCCGCAUGG AGCCCGUGCCCGGCCUGGCCUGGCUGGCCUCCACCGUGAACCUGGAGUUCCAG CACGCCUCCCCCCAGCACGCCGGCCUGUACCUGUGCGUGGUGUACGUGGACGA CCACAUCCACGCCUGGGGCCACAUGACCAUCUCCACCGCCGCCCAGUACCGCA ACGCCGUGGUGGAGCAGCACCUGCCCCAGCGCCAGCCCGAGCCCGUGGAGCCC ACCCGCCCCCACGUGCGCGCCUAACUAGUAGUGACUGACUAGGAUCUGGUUACC ACUAAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACU UACACUUACAAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUA AAAAGAAAGUUUCUUCACAUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAC (SEQ ID NO: 16) [0220] In some embodiments, all uridine residues are 1-methyl-pseudouridine. In some embodiments, underlined residues represent 5’ untranslated sequences (SEQ ID NO: 243). In some embodiments, bold residues represent a signal sequence (leader sequence) (SEQ ID NO: 19) to assist expression of the HSV-2 gE fragment. In some embodiments, italicized residues represent 3’ untranslated sequences (SEQ ID NO: 244) and poly adenylation tail (SEQ ID NO: 264). [0221] In another embodiment, a nucleotide sequence of the RNA encoding an HSV-2 gE fragment lacks the 5’ untranslated sequences, the signal sequence, the 3’ untranslated sequences, the poly adenylation tail, or a combination thereof. In some embodiments, the sequence of the HSV-2 gE fragment is as set forth in SEQ ID NO: 27. [0222] In some embodiments, an HSV-2 gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises amino acids 24-405 of gE from HSV-2 (e.g., strain 2.12 or US8) as set forth in the following amino acid sequence: RTSWKRVTSGEDVVLLPAPAGPEERTRAHKLLWAAEPLDACGPLRPSWVALWPPR RVLETVVDAACMRAPEPLAIAYSPPFPAGDEGLYSELAWRDRVAVVNESLVIYGAL ETDSGLYTLSVVGLSDEARQVASVVLVVEPAPVPTPTPDDYDEEDDAGVSERTPVSV 65 11979815v1 P-627574-PC PPPTPPRRPPVAPPTHPRVIPEVSHVRGVTVHMETPEAILFAPGETFGTNVSIHAIAHD DGPYAMDVVWMRFDVPSSCAEMRIYEACLYHPQLPECLSPADAPCAVSSWAYRLA VRSYAGCSRTTPPPRCFAEARMEPVPGLAWLASTVNLEFQHASPQHAGLYLCVVYV DDHIHAWGHMTISTAAQYRNAVVEQHLPQRQPEPVEPTRPHVRA (SEQ ID NO: 17). [0223] In some embodiments, an HSV-2 gE fragment comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 17. In some embodiments, an HSV-2 gE fragment has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 17. [0224] In some embodiments, full length HSV-2 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the following amino acid sequence: MARGAGLVFFVGVWVVSCLAAAPRTSWKRVTSGEDVVLLPAPAERTRAHKLLWA AEPLDACGPLRPSWVALWPPRRVLETVVDAACMRAPEPLAIAYSPPFPAGDEGLYSE LAWRDRVAVVNESLVIYGALETDSGLYTLSVVGLSDEARQVASVVLVVEPAPVPTP TPDDYDEEDDAGVTNARRSAFPPQPPPRRPPVAPPTHPRVIPEVSHVRGVTVHMETL EAILFAPGETFGTNVSIHAIAHDDGPYAMDVVWMRFDVPSSCADMRIYEACLYHPQ LPECLSPADAPCAVSSWAYRLAVRSYAGCSRTTPPPRCFAEARMEPVPGLAWLAST VNLEFQHASPQHAGLYLCVVYVDDHIHAWGHMTISTAAQYRNAVVEQHLPQRQPE PVEPTRPHVRAPHPAPSARGPLRLGAVLGAALLLAALGLSAWACMTCWRRRSWRA VKSRASATGPTYIRVADSELYADWSSDSEGERDGSLWQDPPERPDSPSTNGSGFEILS PTAPSVYPHSEGRKSRRPLTTFGSGSPGRRHSQASYPSVLW (SEQ ID NO: 18). [0225] In some embodiments, an HSV-2 gE comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence SEQ ID NO: 18. In some embodiments, an HSV-2 gE has an amino acid sequence that is identical to the amino acid sequence SEQ ID NO: 18. [0226] In another embodiment, an HSV-2 gE encoded by RNA utilized in the methods and compositions of the present disclosure comprises the amino acid sequences as set forth in any of the following GenBank Accession Numbers: ABU45436.1, ABU45437.1, ABU45438.1, ABU45439.1, ABW83306.1, ABW83308.1, ABW83310.1, ABW83312.1, ABW83314.1, ABW83316.1, ABW83318.1, ABW83320.1, ABW83322.1, ABW83324.1, ABW83326.1, ABW83328.1, ABW83330.1, ABW83332.1, ABW83334.1, ABW83336.1, ABW83338.1, 66 11979815v1 P-627574-PC ABW83340.1, ABW83342.1, ABW83344.1, ABW83346.1, ABW83348.1, ABW83350.1, ABW83352.1, ABW83354.1, ABW83356.1, ABW83358.1, ABW83360.1, ABW83362.1, ABW83364.1, ABW83366.1, ABW83368.1, ABW83370.1, ABW83372.1, ABW83374.1, ABW83376.1, ABW83378.1, ABW83380.1, ABW83382.1, ABW83384.1, ABW83386.1, ABW83388.1, ABW83390.1, ABW83392.1, ABW83394.1, ABW83396.1, ABW83398.1, ABW83400.1, ABZ04069.1, AEV91407.1, AHG54732.1, AKC42830.1, AKC59307.1, AKC59378.1, AKC59449.1, AKC59520.1, AKC59591.1, AMB66104.1, AMB66173.1, AMB66246.1, AMB66465.1, AQZ55756.1, AQZ55827.1, AQZ55898.1, AQZ55969.2, AQZ56040.2, AQZ56111.2, AQZ56182.1, AQZ56253.2, AQZ56324.1, AQZ56395.1, AQZ56466.2, AQZ56537.1, AQZ56608.1, AQZ56679.1, AQZ56750.1, AQZ56821.2, AQZ56892.1, AQZ56963.2, AQZ57034.2, AQZ57105.1, AQZ57176.1, AQZ57247.2, AQZ57318.2, AQZ57389.2, AQZ57460.2, AQZ57531.2, AQZ57602.2, AQZ57673.1, AQZ57744.2, AQZ57815.1, AQZ57886.1, AQZ57957.2, AQZ58028.2, AQZ58099.1, AQZ58170.2, AQZ58241.2, AQZ58312.2, AQZ58383.2, AQZ58454.2, AQZ58525.2, AQZ58596.1, AQZ58667.1, AQZ58738.2, AQZ58809.2, AQZ58880.2, AQZ58951.2, AQZ59022.2, AQZ59093.1, AQZ59164.1, ARO38081.1, ARO38082.1, ARO38083.1, ARO38084.1, ARO38085.1, ARO38086.1, CAB06715.1, P89436.1, P89475.1, or YP_009137220.1. [0227] In another embodiment, an HSV gE fragment encoded by RNA utilized in the methods and compositions of the present disclosure comprises an IgG Fc-binding domain of the gE protein. In another embodiment, the gE domain encoded by RNA utilized in the methods and compositions of the present disclosure is any other gE domain known in the art to mediate binding to IgG Fc. [0228] In another embodiment, an HSV gE protein encoded by RNA utilized in the methods and compositions of the present disclosure comprises a gE domain involved in cell-to-cell spread. [0229] In another embodiment, an HSV gE fragment encoded by RNA fragment utilized in the methods and compositions of the present disclosure comprises an immune evasion domain. In another embodiment, the gE fragment encoded by RNA fragment utilized in the methods and compositions of the present disclosure comprises a portion of an immune evasion domain. [0230] Each RNA encoding HSV-1 gE or HSV-2 gE protein or fragment thereof represents a separate embodiment of the present disclosure. [0231] In another embodiment, an HSV gE protein fragment encoded by RNA utilized in the methods and compositions of the present disclosure is an immunogenic fragment. In another 67 11979815v1 P-627574-PC embodiment, a gE immunoprotective antigen need not be the entire protein. The protective immune response generally involves, in another embodiment, an antibody response. In another embodiment, mutants, sequence conservative variants, and functional conservative variants of gE are useful in methods and compositions of the present disclosure, provided that all such variants retain the required immuno-protective effect. In another embodiment, the immunogenic fragment can comprise an immuno-protective gE antigen from any strain of HSV. In another embodiment, the immunogenic fragment can comprise sequence variants of HSV, as found in infected individuals. [0232] In some embodiments, an RNA of the present disclosure encodes an HSV polypeptide, or fragment thereof, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 1. [0233] In some embodiments, methods of the present disclosure comprise administering to a subject an HSV polypeptide, or fragment thereof, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 1. Table 1: Exemplary amino acid sequences of HSV immunogens Sequence Amino acid sequence SEQ ID NO:
Figure imgf000070_0001
11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name
Figure imgf000071_0001
69 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name
Figure imgf000072_0001
70 11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name
Figure imgf000073_0001
11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name
Figure imgf000074_0001
11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name
Figure imgf000075_0001
11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name
Figure imgf000076_0001
11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name
Figure imgf000077_0001
11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name
Figure imgf000078_0001
11979815v1 P-627574-PC Sequence Amino acid sequence SEQ ID NO: Name [ ence a
Figure imgf000079_0001
, , , , , , %, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 2. [0235] In some embodiments, methods of the present disclosure comprise administering to a subject an RNA comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 2. Table 2: Exemplary nucleic acid sequences of HSV immunogens HSV-1 gD AAGUACGCCCUGGCCGACGCCUCCCUGAAGAUGGC 22
Figure imgf000079_0002
11979815v1 P-627574-PC GCCCCCGCCUUCGAGACCGCCGGCACCUACCUGCGC CUGGUGAAGAUCAACGACUGGACCGAGAUCACCCA
Figure imgf000080_0001
11979815v1 P-627574-PC CCGAGAACCAGCGCACCGUGGCCCUGUACUCCCUG AAGAUCGCCGGCUGGCACGGCCCCAAGCCCCCCUAC
Figure imgf000081_0001
11979815v1 P-627574-PC UAACUGGCACAUCCCCAGCAUCCAGGAUGUGGCCC CUCAUCAUUGA
Figure imgf000082_0001
11979815v1 P-627574-PC Version CGCGUCUACCACAUACAGCCUAGUCUUGAGGACCC 2.1 UUUUCAGCCACCGUCUAUCCCCAUUACCGUGUACU
Figure imgf000083_0001
11979815v1 P-627574-PC ACCUCACAAUAGCGUGGUAUCGAAUGGGCGAUAAC UGCGCAAUUCCCAUCACAGUCAUGGAGUACACGGA
Figure imgf000084_0001
11979815v1 P-627574-PC AUGCACGCCCCCGCCUUCGAAACCGCCGGCACCUAC CUGAGACUGGUGAAAAUCAACGAUUGGACCGAAAU
Figure imgf000085_0001
11979815v1 P-627574-PC AGCAUUGGCAUGCUGCCUCGUUUCAUUCCCGAGAA UCAACGGACAGUGGCUCUGUAUUCCCUGAAGAUCG
Figure imgf000086_0001
11979815v1 P-627574-PC GCCAGAUCGACACCCAGACCCACGAGCACCCCGACG GCUUCACCACCGUGUCCACCGUGACCUCCGAGGCC
Figure imgf000087_0001
11979815v1 P-627574-PC AGGCCAGCCUUUCAAGGCCACAUGUACCGCCGCCA CCUACUAUCCCGGAAACAGAGCCGAGUUCGUUUGG
Figure imgf000088_0001
11979815v1 P-627574-PC GCACCGACCGCCCCUCCGCCUACGGCACCUGGGUGC GCGUGCGCGUGUUCCGCCCCCCCUCCCUGACCAUCC
Figure imgf000089_0001
11979815v1 P-627574-PC UUGGCCCGCUUGGACGACAGAGACUCAUCAUCGAG GAACUGACACUGGAGACACAGGGGAUGUACUACUG
Figure imgf000090_0001
11979815v1 P-627574-PC CAGGAGGACAGCUGGUCUAUGACUCAGCGCCCAAU AGGACCGAUCCCCACGUGAUCUGGGCAGAAGGAGC
Figure imgf000091_0001
11979815v1 P-627574-PC UGAGUUUCGGCUGCAGAUCUGGCGUUAUGCCACAG CUACUGACGCAGAGAUUGGUACCGCCCCCAGUUUG
Figure imgf000092_0001
11979815v1 P-627574-PC CCCAAAGACAAGUAGCGAACCAGUUCGGUGCAACA GGCAUGACCCACUUGCACGCUAUGGGUCAAGAGUC
Figure imgf000093_0001
11979815v1 P-627574-PC (28-426) GGAAUGCUUCUGCCCCAAGAACUACCCCCACUCCU Version CCUCAACCCAGGAAAGCGACAAAGUCCAAGGCCAG
Figure imgf000094_0001
92 11979815v1 P-627574-PC UGGCCGGCUAUCCCGAUGGGAUUCCAGUCCUGGAG CACCACUGA
Figure imgf000095_0001
11979815v1 P-627574-PC AGAAAGUGGCUGUGGCCUCUCAGACGAGCUGCGGU CGACCAGGAACAGCUACCAUUCGCAGCACUCUGCC
Figure imgf000096_0001
11979815v1 P-627574-PC CCAUGGAAUUCACCGGCGAUCACGCCGUGUGCACC GCCGGCUGCGUGCCCGAAGGCGUGACCUUCGCCUG
Figure imgf000097_0001
11979815v1 P-627574-PC CCUUCACCUGCCAGCUGACCUGGCACCGCGACUCCG UGUCCUUCUCCCGCCGCAACGCCUCCGGCACCGCCU
Figure imgf000098_0001
11979815v1 P-627574-PC ACACAGACCCAAGAGAACCCCGACGGCUUUAGCAC CGUGUCCACAGUGACAUCUGCCGCCGUUGGAGGAC
Figure imgf000099_0001
11979815v1 P-627574-PC CCUUUCAAGGCCACAUGUACCGCCGCCACCUACUA UCCCGGAAACAGAGCCGAGUUCGUUUGGUUCGAGG
Figure imgf000100_0001
11979815v1 P-627574-PC UGGGGCCGUACUGACCGCCCUUCCGCAUAUGGCAC UUGGGUGAGAGUUCGCGUCUUUCGGCCCCCUUCUC
Figure imgf000101_0001
11979815v1 P-627574-PC CCUGGGGCCUCUCCACGGCUGUACUCAGUUGUUGG CCCGCUUGGACGACAGAGACUCAUCAUCGAAGAGC
Figure imgf000102_0001
11979815v1 P-627574-PC AGAAGUGAUGGUGAACGUGUCCGCCCCUCCUGGCG GCCAGCUGGUGUACGAUUCCGCCCCCAACAGAACC
Figure imgf000103_0001
11979815v1 P-627574-PC GCAGAUCAGAUGCAGAUUUCCAAAUUCCACCAGAA CAGAAUUCAGACUCCAGAUCUGGAGAUAUGCAACA
Figure imgf000104_0001
11979815v1 P-627574-PC CGGCCAAACCCGCCCCUCCACCCAAAACCGGACCUC CUAAGACCAGCUCUGAACCGGUGCGGUGUAAUAGG
Figure imgf000105_0001
103 11979815v1 P-627574-PC HSV-2 gC AGUCCAGGAAGGACGAUUACGGUGGGACCCAGAGG 130 [UL44] UAAUGCGUCCAAUGCUGCGCCAUCCGCUUCUCCAC
Figure imgf000106_0001
104 11979815v1 P-627574-PC GUUUCCUACGAACAGACCGAGUAUAUCUGCCGAUU GGCCGGUUACCCCGAUGGGAUACCAGUCCUGGAGC
Figure imgf000107_0001
11979815v1 P-627574-PC CGCCUGGUUUCUCGGUGAUGACUCCUCUCCUGCUG AAAAGGUGGCUGUAGCCUCCCAAACAAGCUGUGGU
Figure imgf000108_0001
11979815v1 P-627574-PC UGCGUCAGGGACCGCCUCCGUGCUUCCUCGGCCAA CCAUCACAAUGGAAUUCACGGGUGAUCACGCUGUC
Figure imgf000109_0001
11979815v1 P-627574-PC CGUGUCAACGGUCACAUCUGCCGCCGUCGGAGGAC AAGGGCCACCCAGAACCUUCACAUGCCAGCUGACC
Figure imgf000110_0001
11979815v1 P-627574-PC UGGACGUGGUGUGGCUGCGCUUCGACGUGCCCACC UCCUGCGCCGAGAUGCGCAUCUACGAGUCCUGCCU
Figure imgf000111_0001
11979815v1 P-627574-PC CGCCCACGACGACGGCCCCUACGCCAUGGACGUGG UGUGGAUGCGCUUCGACGUGCCCUCCUCCUGCGCC
Figure imgf000112_0001
11979815v1 P-627574-PC CUGGCGAGACAUUUGGCACCAACGUGUCCAUCCAC GCUAUCGCCCACGACGAUGGCCCUUACGCCAUGGA
Figure imgf000113_0001
11979815v1 P-627574-PC GCACAUGGAAACACCUGAGGCCAUCCUGUUCGCCC CUGGCGAGACAUUUGGCACCAACGUGUCCAUCCAC
Figure imgf000114_0001
11979815v1 P-627574-PC AUUCCCGAGGUCAGCCAUGUGCGCGGCGUAACUGU GCACAUGGAGACGCCCGAAGCGAUACUGUUUGCCC
Figure imgf000115_0001
11979815v1 P-627574-PC GACCCCCAGUAGCACCUCCAACCCAUCCGAGAGUG AUUCCCGAGGUCAGCCAUGUGCGCGGCGUAACUGU
Figure imgf000116_0001
11979815v1 P-627574-PC ACCGGUGAGUGUGCCACCUCCCACACCGCCAAGGA GACCCCCAGUAGCACCUCCAACCCAUCCGAGAGUG
Figure imgf000117_0001
11979815v1 P-627574-PC UGAAGAAGAUGAUGCCGGCGUGUCCGAAAGAACCC CCGUGUCCGUGCCCCCUCCCACCCCUCCCCGCAGAC
Figure imgf000118_0001
11979815v1 P-627574-PC CCAGCUCCAGUGCCAACCCCAACCCCAGAUGAUUA UGAUGAAGAAGAUGAUGCUGGAGUGUCUGAAAGA
Figure imgf000119_0001
11979815v1 P-627574-PC ACGGCAAGUGGCUUCCGUGGUACUGGUCGUAGAGC CCGCACCAGUUCCCACACCCACCCCGGAUGACUACG
Figure imgf000120_0001
11979815v1 P-627574-PC CCUCCCUGCCCAUCACCGUGUACUACGCCGUGCUG GAGCGCGCCUGCCGCUCCGUGCUGCUGAACGCCCCC
Figure imgf000121_0001
11979815v1 P-627574-PC AAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUA GCAUGACCCGCCUGACCGUGCUGGCCCUGCUGGC
Figure imgf000122_0001
0 11979815v1 P-627574-PC UUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUCAC AUUCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
Figure imgf000123_0001
11979815v1 P-627574-PC ACCUGCACCGCCGCCGCCUACUACCCCCGCAACCCC GUGGAGUUCGACUGGUUCGAGGACGACCGCCAGGU
Figure imgf000124_0001
11979815v1 P-627574-PC GCCCGCCCCCCCCCCCAAGACCGGCCCCCCCAAGAC CUCCUCCGAGCCCGUGCGCUGCAACCGCCACGACCC
Figure imgf000125_0001
3 11979815v1 P-627574-PC CUUACAAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGU AUCUGCUCCUAAUAAAAAGAAAGUUUCUUCACAUUCU
Figure imgf000126_0001
11979815v1 P-627574-PC CCGAGAUGCGCAUCUACGAGUCCUGCCUGUACCAC CCCCAGCUGCCCGAGUGCCUGUCCCCCGCCGACGCC
Figure imgf000127_0001
11979815v1 P-627574-PC GGCCUGUACUCCGAGCUGGCCUGGCGCGACCGCGU GGCCGUGGUGAACGAGUCCCUGGUGAUCUACGGCG
Figure imgf000128_0001
126 11979815v1 P-627574-PC AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAC [ oded
Figure imgf000129_0001
of a sequence provided herein. In another embodiment, an HSV glycoprotein, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure is an isoform of the sequence provided herein. In another embodiment, an HSV glycoprotein, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure is a variant of the sequence provided herein. In another embodiment, an HSV glycoprotein, or immunogenic fragment thereof, encoded by RNA utilized in the methods and compositions of the present disclosure is a fragment of the sequence provided herein. [0237] In another embodiment, a glycoprotein fragment encoded by RNA of the methods and compositions of the present disclosure comprises the ectodomain of the glycoprotein. In another embodiment, a glycoprotein fragment encoded by RNA of the methods and compositions of the present disclosure consists of the ectodomain of the glycoprotein. In another embodiment, a glycoprotein fragment encoded by RNA of the methods and compositions of the present disclosure comprises a fragment of the ectodomain of the glycoprotein. In another embodiment, the glycoprotein fragment may be any glycoprotein fragment known in the art. [0238] In another embodiment, a glycoprotein or immunogenic fragment encoded by an RNA utilized in the methods and compositions of the present disclosure may be from any strain of HSV. In another embodiment, an immunogenic fragment encoded by RNA utilized in the methods and compositions of the present disclosure may comprise sequence variants of HSV, as found in infected individuals. [0239] In some embodiments, “variant” refers to an amino acid or nucleic acid sequence (or in other embodiments, an organism or tissue) that is different from the majority of the population but is still sufficiently similar to the common mode to be considered to be one of them, for example splice variants. In some embodiments, the variant may a sequence conservative variant, while in another embodiment, the variant may be a functional conservative variant. In some embodiments, a variant may comprise an addition, deletion or substitution of one or more amino acids. [0240] “Immune evasion domain” refers, in some embodiments, to a domain that interferes with or reduces in vivo anti-HSV efficacy of anti-HSV antibodies (e.g., anti-gD antibodies). In another 127 11979815v1 P-627574-PC embodiment, the domain interferes or reduces in vivo anti-HSV efficacy of an anti-HSV immune response. In another embodiment, the domain reduces the immunogenicity of an HSV protein (e.g. gD) during subsequent infection. In another embodiment, the domain reduces the immunogenicity of an HSV protein during subsequent challenge. In another embodiment, the domain reduces the immunogenicity of HSV during subsequent challenge. In another embodiment, the domain reduces the immunogenicity of an HSV protein in the context of ongoing HSV infection. In another embodiment, the domain reduces the immunogenicity of HSV in the context of ongoing HSV infection. In another embodiment, the domain functions as an IgG Fc receptor. In another embodiment, the domain promotes antibody bipolar bridging, which in some embodiments, is a term that refers to an antibody molecule binding by its Fab domain to an HSV antigen and by its Fc domain to a separate HSV antigen, such as in some embodiments, gE, thereby blocking the ability of the Fc domain to activate complement. [0241] The present disclosure also provides for modified RNA encoding analogs of HSV proteins or polypeptides, or fragments thereof. Analogs may differ from naturally occurring proteins or peptides by conservative amino acid sequence substitutions or by modifications which do not affect sequence, or by both. [0242] In another embodiment, an HSV glycoprotein, or an immunogenic fragment thereof, encoded by modified RNA of the present disclosure is homologous to a sequence set forth hereinabove, either expressly or by reference to a GenBank entry. The terms “homology,” “homologous,” etc., when in reference to any protein or peptide, refer, in some embodiments, to a percentage of amino acid residues in the candidate sequence that are identical with the residues of a corresponding native polypeptide, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent homology, and not considering any conservative substitutions as part of the sequence identity. Methods and computer programs for the alignment are well known in the art. [0243] In another embodiment, “homology” refers to identity of a protein sequence encoded by an RNA to a sequence disclosed herein of greater than 70%. In another embodiment, the identity is greater than 72%. In another embodiment, the identity is greater than 75%. In another embodiment, the identity is greater than 78%. In another embodiment, the identity is greater than 80%. In another embodiment, the identity is greater than 82%. In another embodiment, the identity is greater than 83%. In another embodiment, the identity is greater than 85%. In another embodiment, the identity is greater than 87%. In another embodiment, the identity is greater than 128 11979815v1 P-627574-PC 88%. In another embodiment, the identity is greater than 90%. In another embodiment, the identity is greater than 92%. In another embodiment, the identity is greater than 93%. In another embodiment, the identity is greater than 95%. In another embodiment, the identity is greater than 96%. In another embodiment, the identity is greater than 97%. In another embodiment, the identity is greater than 98%. In another embodiment, the identity is greater than 99%. In another embodiment, the identity is 100%. [0244] In some embodiments, “isoform” refers to a version of a molecule, for example, a protein, with only slight differences to another isoform of the same protein. In some embodiments, isoforms may be produced from different but related genes, or in another embodiment, may arise from the same gene by alternative splicing. In another embodiment, isoforms are caused by single nucleotide polymorphisms. [0245] In another embodiment, the RNA encoding a glycoprotein or glycoprotein fragment as described herein further encodes an antigenic tag. In some embodiments, the tag is a histidine (“His”) tag. In some embodiments, the His tag comprises 5 histidine residues. In another embodiment, the His tag comprises 6 histidine residues. [0246] In another embodiment, methods and compositions of the present disclosure utilize a chimeric molecule, comprising a fusion of an RNA encoding an HSV protein, or an immunogenic fragment thereof, with an RNA encoding a tag polypeptide that provides an epitope to which an anti-tag antibody can selectively bind. The epitope tag is placed, in other embodiments, at the amino- or carboxyl-terminus of the protein or in an internal location therein. The presence of such epitope-tagged forms of the recombinant HSV protein, or an immunogenic fragment thereof, is detected, in another embodiment, using an antibody against the tag polypeptide. In another embodiment, inclusion of the epitope tag enables the recombinant HSV protein, or an immunogenic fragment thereof, to be readily purified by affinity purification using an anti-tag antibody or another type of affinity matrix that binds to the epitope tag. Various tag polypeptides and their respective antibodies are known in the art. [0247] In some embodiments, the compositions of the present disclosure comprise an adjuvant, while in another embodiment, the compositions do not comprise an adjuvant. “Adjuvant” refers, in another embodiment, to compounds that, when administered to an individual or tested in vitro, increase the immune response to an antigen in the individual or test system to which the antigen is administered. In another embodiment, an immune adjuvant enhances an immune response to an antigen that is weakly immunogenic when administered alone, i.e., inducing no or weak antibody 129 11979815v1 P-627574-PC titers or cell-mediated immune response. In another embodiment, the adjuvant increases antibody titers to the antigen. In another embodiment, the adjuvant lowers the dose of the antigen effective to achieve an immune response in the individual. Multiple types of adjuvants are known in the art and described in detail in U. S. Patent Publication 2013/0028925 which is hereby incorporated by reference herein. Signal Sequence and Signal Peptides [0248] In some embodiments, an RNA encoding a glycoprotein as described herein comprises a signal sequence encoding a signal peptide, e.g., that is functional in mammalian cells. In some embodiments, a signal sequence encodes a modified signal peptide (e.g., comprising amino acid substitutions or amino acid additions). In some embodiments, a signal sequence is a codon optimized signal sequence. [0249] In some embodiments, a utilized signal sequence is a heterologous signal sequence. In some embodiments, a heterologous signal sequence comprises or consists of a non-human signal sequence. In some embodiments, a heterologous signal sequence comprises or consists of a viral signal sequence. In some embodiments, a viral signal sequence comprises or consists of an HSV signal sequence (e.g., an HSV-1 or HSV-2 signal sequence). In some embodiments, a signal sequence comprises or consists of an HSV-1 signal sequence. In some embodiments, a signal sequence comprises or consists of an HSV-2 signal sequence. In some embodiments, a signal sequence is characterized by a length of about 15 to 30 amino acids. In some embodiments, a signal sequence encodes a signal peptide that preferably allows transport of an HSV-1 glycoprotein, an HSV-2 glycoprotein, an HSV-1 and HSV-2 glycoprotein, or an immunogenic fragment of an HSV-1 and/or HSV-2 glycoprotein, with which it is associated into a defined cellular compartment, preferably a cell surface, endoplasmic reticulum (ER) or endosomal- lysosomal compartment. [0250] In some embodiments, a signal sequence is the native signal sequence of the encoded glycoprotein. In some embodiments, a signal sequence is or comprises an HSV glycoprotein D (gD) signal sequence (e.g., an HSV-1 or HSV-2 gD signal sequence). In some embodiments, a signal peptide encoded by a signal sequence is or comprises an HSV-2 gD signal peptide (SEQ ID NO: 29). In another embodiment, an HSV-2 gD signal peptide comprises KY (SEQ ID NO: 30), KYA (SEQ ID NO: 31, KYAL (SEQ ID NO: 32), or KYALA (SEQ ID NO: 33) at the C terminus of the signal peptide. In some embodiments, a signal peptide is or comprises an HSV-1 gD signal 130 11979815v1 P-627574-PC peptide (SEQ ID NO: 34). In some embodiments, an HSV-1 gD signal peptide comprises KY at the C terminus of the signal sequence (SEQ ID NO: 35). [0251] In some embodiments, a signal sequence is or comprises an HSV glycoprotein C (gC) signal sequence (e.g., an HSV-1 or HSV-2 gC signal sequence). In another embodiment, a signal peptide encoded by a signal sequence is or comprises an HSV-2 gC signal peptide (SEQ ID NO: 36). In another embodiment, a signal peptide is or comprises an HSV-1 gC signal peptide (SEQ ID NO: 37). [0252] In some embodiments, a signal sequence is or comprises an HSV glycoprotein E (gE) signal sequence (e.g., an HSV-1 or HSV-2 gE signal sequence). In some embodiments, a signal peptide encoded by a signal sequence is or comprises an HSV-2 gE signal sequence (SEQ ID NO: 38). In some embodiments, a signal peptide is or comprises an HSV-1 gE signal peptide (SEQ ID NO: 39). In some embodiments, an HSV-2 gE signal peptide comprises RTS at the C terminus of the signal peptide (SEQ ID NO: 40). In some embodiments, an HSV-2 gE signal peptide comprises A20V, A21V, and A22V substitutions (SEQ ID NO: 41). [0253] In some embodiments, a signal sequence is or comprises an HSV gB signal sequence (e.g., an HSV-1 or HSV-2 gB signal sequence). In some embodiments, a signal peptide encoded by a signal sequence is or comprises an HSV-1 gB signal peptide (SEQ ID NO: 42). In some embodiments, a signal peptide encoded by a signal sequence is or comprises an HSV-2 gB signal peptide (SEQ ID NO: 43). In some embodiments, an HSV-1 gB signal peptide comprises AP at the C terminus of the signal peptide (SEQ ID NO: 44). [0254] In some embodiments, a signal sequence is or comprises an HSV gI signal sequence (e.g., an HSV-1 or HSV-2 gI signal sequence). In some embodiments, a signal peptide encoded by a signal sequence is or comprises an HSV-1 gI signal peptide (SEQ ID NO: 45). In some embodiments, an HSV-2 gI signal sequence is or comprises an HSV-2 gI signal peptide (SEQ ID NO: 249).In some embodiments, a signal peptide encoded by a signal sequence is or comprises amino acid residues 1-18 of the wild-type sequence (SEQ ID NO: 46). In some embodiments, an HSV-2 gI signal peptide comprises an additional leucine residue at the C terminus of the signal peptide (SEQ ID NO: 47). [0255] In other embodiments, a signal sequence is a heterologous signal sequence. In some embodiments, a heterologous signal peptide encoded by a heterologous signal sequence comprises an IL-2 signal peptide (SEQ ID NO: 220). In some embodiments, a heterologous signal peptide encoded by a heterologous signal sequence comprises or consists of an azurocidin signal peptide 131 11979815v1 P-627574-PC (SEQ ID NO: 221). In some embodiments, a heterologous signal peptide encoded by a heterologous signal sequence comprises or consists of an MHC Class II signal peptide (SEQ ID NO: 222). In some embodiments, a signal sequence comprises or consists of an Ebola virus signal sequence. In some embodiments, an Ebola virus signal peptide comprises or consists of an Ebola virus spike glycoprotein (SGP) signal peptide (SEQ ID NO: 48). In other embodiments, an RNA encoding a glycoprotein as described herein does not comprise a signal sequence. In some embodiments, an RNA as described herein comprises only an ectodomain. [0256] In some embodiments, a signal sequence encodes an exemplary signal peptide listed in Table 3, or a signal peptide having 1, 2, 3, 4, or 5 amino acid differences relative thereto. In some embodiments, a signal peptide is an exemplary signal peptide selected from those listed in Table 3 and functionally connected to the N-terminus of an exemplary HSV immunogen selected from those listed in Table 1. Table 3: Exemplary signal peptides Sequence Name Amino Acid Sequence SEQ ID NO:
Figure imgf000134_0001
11979815v1 P-627574-PC Sequence Name Amino Acid Sequence SEQ ID NO: HSV-2 gE Signal Sequence MARGAGLVFFVGVWVVSCLVVVP 41 [ ed in T
Figure imgf000135_0001
able 4, or a signal sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence listed in Table 4. In some embodiments, an exemplary signal sequence is selected from those listed in Table 4 and functionally connected (i.e., in frame) to the 5' end of an exemplary HSV immunogen nucleic acid sequence selected from those listed in Table 2. 133 11979815v1 P-627574-PC Table 4: Exemplary signal sequences Sequence Name Version Amino Acid Sequence SEQ ID NO: HSV-2 gD WT AUGGGCCGCCUGACCUCCGGCGUGGGC 53
Figure imgf000136_0001
11979815v1 P-627574-PC Sequence Name Version Amino Acid Sequence SEQ ID NO: UUGGUCUGAGAGUUGUGUGUGCCAAA
Figure imgf000137_0001
11979815v1 P-627574-PC Sequence Name Version Amino Acid Sequence SEQ ID NO: HSV-1 gD Version 4 AUGGGCGGCGCAGCCGCUAGACUGGG 255
Figure imgf000138_0001
136 11979815v1 P-627574-PC Sequence Name Version Amino Acid Sequence SEQ ID NO: HSV-2 gC Version 2 AUGGCCUUGGGGAGAGUGGGCCUUGC 71
Figure imgf000139_0001
11979815v1 P-627574-PC Sequence Name Version Amino Acid Sequence SEQ ID NO: HSV-2 gE RTS Version 2 AUGGCGAGAGGAGCCGGGCUCGUGUU 80
Figure imgf000140_0001
138 11979815v1 P-627574-PC Sequence Name Version Amino Acid Sequence SEQ ID NO: UGCUGGGACUGACACUGGGAGUUCUG
Figure imgf000141_0001
139 11979815v1 P-627574-PC Sequence Name Version Amino Acid Sequence SEQ ID NO: HSV-2 gI L Version 2 AUGCCCGGCAGAAGCCUCCAGGGACU 89
Figure imgf000142_0001
140 11979815v1 P-627574-PC Sequence Name Version Amino Acid Sequence SEQ ID NO: IL2 Version 3 AUGAGAAUGCAGCUGCUGCUGCUGAU 52
Figure imgf000143_0001
[0258] The present invention also provides RNA comprising a nucleotide sequence encoding a protein, wherein the protein comprises an HSV (e.g., HSV-1, HSV-2, or both) glycoprotein antigen or immunogenic fragment thereof and a signal peptide. In some embodiments, a nucleotide sequence encodes a protein, wherein the protein comprises an HSV-2 glycoprotein antigen, or immunogenic fragment thereof, and a signal peptide. 141 11979815v1 P-627574-PC [0259] Exemplary proteins comprising signal peptides and HSV-2 immunogens are shown in Table 5. In some embodiments, a protein of the present disclosure comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 5. Table 5. Exemplary signal peptide/HSV-2 immunogen combinations HSV SEQ Signal Glycoprotei Amino Acid Sequence S P R N R R T P S E N A P S Y S D S
Figure imgf000144_0001
142 11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence T K G S E Q A C H S S L G I T G H
Figure imgf000145_0001
143 11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence R A W M F G P T R A Y P C H S E G L
Figure imgf000146_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence I I G H T T A I P P E I G H T T A I T R A Y
Figure imgf000147_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence P C H S E R A W M F G P T K G S E Q
Figure imgf000148_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence A C H TI T Y E Y E Q A C H P H D I
Figure imgf000149_0001
147 11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence P R T D T G N A S T T E S V TI G T
Figure imgf000150_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence I G K T R G T V S S L G I T G H T K G S E
Figure imgf000151_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence Q A C H S P P E I G H T T A I A Q D A H P E
Figure imgf000152_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence G H V P Y E N P A S P P A P A P H K N
Figure imgf000153_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence A S S I Y F A F T IP K E E S I T IP L A L E S
Figure imgf000154_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence D P L G Y V A C N D P M L
Figure imgf000155_0001
153 11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence S E I A V M L G R P S L R V G R P
Figure imgf000156_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence S L R V M P A S R S T R C A L T V A C
Figure imgf000157_0001
11979815v1 P-627574-PC HSV SEQ Signal Glycoprotei Amino Acid Sequence N
Figure imgf000158_0001
[0260] The present disclosure also provides RNAs comprising nucleotide sequences as provided herein. In some embodiments, an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gC protein or immunogenic fragment thereof. In some embodiments, an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gD protein or immunogenic fragment thereof. In some embodiments, an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gE protein or immunogenic fragment thereof. [0261] Exemplary RNA comprising a signal sequence (i.e., encoding a signal peptide) and a sequence encoding an HSV-2 glycoprotein are shown in Table 6 below. In some embodiments, an RNA of the present disclosure comprises a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6. Table 6: Exemplary signal sequence/HSV-2 immunogen nucleotide sequence combinations SEQ Signal Immunoge Versio Nucleotide Sequence C C G A
Figure imgf000158_0002
156 11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C U U A A C A G C C G C G C G G C G C C G C C C
Figure imgf000159_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C C A C U U C A G A A C C C U G G G U C C C U
Figure imgf000160_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C C A G U U G A A A C A G A G A A C A A A A C
Figure imgf000161_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A G A U A G G A C C A G A C U C A C G C C A
Figure imgf000162_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G G C C C C C A C C C C A G C A C C A C G G G
Figure imgf000163_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G G G C C C C C G C C C C C G C C C G C C C
Figure imgf000164_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C G C G G C C G G C G G C C C C G C G C G C C G G C
Figure imgf000165_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C U C G A G C A A C A G A G U A C C U C
Figure imgf000166_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C A G U A G A C A C A G A U C G A G C A A
Figure imgf000167_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C A G A G U A C C U C C C A G U A G A C A C
Figure imgf000168_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A G A U C G A G C A A C A G A G U A C C U
Figure imgf000169_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C C A G U A G A C A C A G A C U C A C U C
Figure imgf000170_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C G C G A A U G C U G G A A G A G A A U C C
Figure imgf000171_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C C A U C U A A A C G U C G A G C C G C
Figure imgf000172_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U G G A A G A G A A U A C A C A U C U A A A C
Figure imgf000173_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G U C G A G A G G C U G G A A G A G A A U A
Figure imgf000174_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C A C A C G C C C A C C C C A G C A C C A C G
Figure imgf000175_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G G G G G C C C C C G C C C C C G C C G A C A U
Figure imgf000176_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A U C A C C C C G U A U G U C U G A U G G G
Figure imgf000177_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U A A A U G A C A U A U C A C C C C
Figure imgf000178_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G U A U G U C U G A U G G G U A A A G A C A
Figure imgf000179_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U A U C A C C C C G U A U G U C U G A U G G
Figure imgf000180_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G U A A A G C C C C C G A U A C C C C
Figure imgf000181_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G U A U G U C U G A U G G G U A A A C U A
Figure imgf000182_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C U A C G C A A C G U U U G C A C A G A C G G C U C
Figure imgf000183_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A A A G G U C G C C A C A C A G C C A C
Figure imgf000184_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C G G U A G G A G C U C C U G U C C C
Figure imgf000185_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C C C A G U U G G A U U C U
Figure imgf000186_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U A U G A A U G C C A C G A A U C
Figure imgf000187_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A G C G G U A G A U G A C G U C A C U A
Figure imgf000188_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C U G C A G A G C U G U C G C C G G G C U A A
Figure imgf000189_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C U C G G G C G A C U C C C G G A C G G C C G
Figure imgf000190_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C C U C C A C U G U U G A G G C C U U G
Figure imgf000191_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A C A C C U C U U C C C U U C A U
Figure imgf000192_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U C G C A U A C U G C C C A C G U G
Figure imgf000193_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C G C G G A G G C A A G C A C G U G U G U
Figure imgf000194_0001
192 11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C G C C G C C G G C A C A G A A U G C U G G A A
Figure imgf000195_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G A G A A U C C C C C A U U G U A G A C C G U C G
Figure imgf000196_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A G C C G C U G G A A G A G A A U A C A C A
Figure imgf000197_0001
195 11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A G C C G C C G C G C G U C U A G
Figure imgf000198_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A C A G A G U A G C C G C C G G C G G
Figure imgf000199_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A U C U G C U U A U C U C A G C C C U U
Figure imgf000200_0001
198 11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A G A A A C U C A A G G G U C G U G U U C A G A
Figure imgf000201_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C U A C G G C G A C A A A G C G U U C G G C
Figure imgf000202_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C C G A G C A C U G A G C G C C C G C A
Figure imgf000203_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U A A A A U C A C C C A G A C A G A
Figure imgf000204_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C U U A C A C A U A G A C C G G C
Figure imgf000205_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C C G A G C A C U G U A G C G C C A U G
Figure imgf000206_0001
204 11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U C C C U C A C G C C U C G U A U A G C A C
Figure imgf000207_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U C A U C G A C G C C U C U C U G C G A C G
Figure imgf000208_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G A C C A U C G U A U A U C A C C A A G C G A G
Figure imgf000209_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C U A A U U A A A G A A A C A
Figure imgf000210_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C U G C A A C C U C U A C C C A C
Figure imgf000211_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C G U A C C C C U A C U U C A C C U C U U
Figure imgf000212_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G U A C G C C G G G C C C G C C C G C C G G C G C
Figure imgf000213_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C G G C C C C C U G G C G A U C C G G A G U U
Figure imgf000214_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C A C A G C C C U A G C G U U A C U C C A U C A
Figure imgf000215_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U G C A U U C C G U U C U U A C U C C
Figure imgf000216_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A U C A U G C A U U C C G U U C U C G C C
Figure imgf000217_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G A A U C C G C C C G C C G C U C A C C A A A C A
Figure imgf000218_0001
216 11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A C C G C G C G G G G G G C G C C C C C C G
Figure imgf000219_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A C G C C G G U U C U C A G U G U G U G C G
Figure imgf000220_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G U U U U U G A C G U U U G C G G C A U C C G
Figure imgf000221_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A G U U C A U A U G A C A C C G A C A U A C U
Figure imgf000222_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U G A C C A A C A A C G C G G A A U C A U
Figure imgf000223_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A C C C C U G A C G C C U A G A G C G C G C C C G C G
Figure imgf000224_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C G C G G G C G A C A G G A C C C C C G G C
Figure imgf000225_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C G C G U G U G U G C G G U U U U
Figure imgf000226_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U G A C G U U U G C G G C A U U G C C G U G C C
Figure imgf000227_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C G C G A C C U U C U A C U U G A C C A
Figure imgf000228_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A U G C C G U G C C C G C G A C C U U C U
Figure imgf000229_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A C U U G A C C A A C G C C U G G U C U G
Figure imgf000230_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C C U U C U A A C C C A G G U C G C G
Figure imgf000231_0001
229 11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G G C A C A G C G U G C U C A A A G G G C G U
Figure imgf000232_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C G U U G C C G G C A C A G C G U G C U C
Figure imgf000233_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A A A G G G C G U G C G U U G C C G G C A
Figure imgf000234_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C A G C G U G C U C A A A G G G C G U G C G U
Figure imgf000235_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n U G C C G G C A C A G C G U G C U C A A A G
Figure imgf000236_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G G C G U G C G U U G C C G G C A C A G C
Figure imgf000237_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G U G C U C A A A G G G C G U G C G U U G C C
Figure imgf000238_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G C C C U C G A C C G C A U U C U A A U A G
Figure imgf000239_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G G C U C C C C C G A U A C U C A
Figure imgf000240_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C C A A C C C A C G A U G C G G C C G U C U
Figure imgf000241_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A G G U C C G C G A A A C C G A C U C C
Figure imgf000242_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A G A C C U A U C C G U G C C G
Figure imgf000243_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n A C U G U U C A G A C C U A G
Figure imgf000244_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n G U C C G C G A A A C C G A C U C C A G A
Figure imgf000245_0001
11979815v1 P-627574-PC SEQ Signal Immunoge Versio Nucleotide Sequence ID Sequenc n n C C U A U
Figure imgf000246_0001
[0262] In some embodiments, an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gC protein or immunogenic fragment thereof and a signal sequence. In some embodiments, an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gD protein or immunogenic fragment thereof and a signal sequence. In some embodiments, an RNA provided herein comprises a nucleotide sequence that encodes an HSV-2 gE protein or immunogenic fragment thereof and a signal sequence. [0263] In some embodiments, nucleotide sequences described herein can comprise a nucleotide sequence that encodes a 5’UTR and/or a 3’ UTR. In some embodiments, polynucleotides described herein can comprise a nucleotide sequence that encodes a polyA tail. In some embodiments, nucleotide sequences described herein may comprise a 5’ cap, which may be incorporated during transcription, or joined to a nucleotide sequence post-transcription. [0264] In some embodiments, a nucleotide sequence provided herein encodes one or more glycoproteins (e.g., glycoprotein C (gC), glycoprotein D (gD), glycoprotein E (gE), or a combination thereof), or an immunogenic fragment thereof. In some embodiments, an RNA comprises a 5’ cap, a 5’UTR, a nucleotide sequence that encodes one or more glycoproteins (e.g., glycoprotein C (gC), glycoprotein D (gD), glycoprotein E (gE), or a combination thereof), or an immunogenic fragment thereof, a 3’ UTR, and a polyA tail. 1. 5' Cap [0265] A structural feature of messenger RNA (mRNA) is a cap structure at the five-prime end (5'). Natural eukaryotic mRNA comprise a 7-methylguanosine cap linked to the mRNA via a 5´ to 5´-triphosphate bridge resulting in a cap0 structure (m7GpppN). In most eukaryotic mRNA and 244 11979815v1 P-627574-PC some viral mRNA, further modifications can occur at the 2'-hydroxyl-group (2’-OH) (e.g., the 2'- hydroxyl group may be methylated to form 2'-O-Me) of the first and subsequent nucleotides producing “cap1” and “cap2” five-prime ends, respectively). Diamond, et al., (2014) Cytokine & growth Factor Reviews, 25:543–550, which is incorporated herein by reference in its entirety, reported that cap0-mRNA cannot be translated as efficiently as cap1-mRNA in which the role of 2'-O-Me in the penultimate position at the mRNA 5’ end is determinant. Lack of the 2'-O-met has been shown to trigger innate immunity and activate an interferon (IFN) response. Daffis, et al. (2010) Nature, 468:452-456; and Züst et al. (2011) Nature Immunology, 12:137-143, each of which is incorporated herein by reference in its entirety. [0266] RNA capping is well researched and is described, e.g., in Decroly E et al. (2012) Nature Reviews 10: 51-65; and in Ramanathan A. et al., (2016) Nucleic Acids Res; 44(16): 7511–7526, the entire contents of each of which are hereby incorporated by reference. For example, in some embodiments, a 5’-cap structure which may be suitable in the context of the present invention is a cap0 (methylation of the first nucleobase, e.g., m7GpppN), cap1 (additional methylation of the ribose of the adjacent nucleotide of m7GpppN), cap2 (additional methylation of the ribose of the 2nd nucleotide downstream of the m7GpppN), cap3 (additional methylation of the ribose of the 3rd nucleotide downstream of the m7GpppN), cap4 (additional methylation of the ribose of the 4th nucleotide downstream of the m7GpppN), ARCA (“anti-reverse cap analogue”), modified ARCA (e.g. phosphothioate modified ARCA), inosine, N1 -methyl-guanosine, 2’-fluoro- guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2- azido-guanosine. [0267] The term “5'-cap” as used herein refers to a structure found on the 5'-end of an RNA, e.g., mRNA, and generally includes a guanosine nucleotide connected to an RNA, e.g., mRNA, via a 5'- to 5'-triphosphate linkage (also referred to as Gppp or G(5')ppp(5')). In some embodiments, a guanosine nucleoside included in a 5’ cap may be modified, for example, by methylation at one or more positions (e.g., at the 7-position) on a base (guanine), and/or by methylation at one or more positions of a ribose. In some embodiments, a guanosine nucleoside included in a 5’ cap comprises a 3’O methylation at a ribose (3’OMeG). In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine (m7G). In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7'-position of guanine and a 3' O methylation at a ribose (m7(3'OMeG)). It will be understood that the notation used in the 245 11979815v1 P-627574-PC above paragraph, e.g., “(m27,3'-O)G” or “m7(3'OMeG)”, applies to other structures described herein. [0268] In some embodiments, providing an RNA with a 5'-cap disclosed herein may be achieved by in vitro transcription, in which a 5'-cap is co-transcriptionally incorporated into an RNA strand, or may be attached to an RNA post-transcriptionally using capping enzymes. In some embodiments, co-transcriptional capping with a cap disclosed herein improves the capping efficiency of an RNA compared to co-transcriptional capping with an appropriate reference comparator. In some embodiments, improving capping efficiency can increase a translation efficiency and/or translation rate of an RNA, and/or increase expression of an encoded protein. In some embodiments, alterations to polynucleotides generate a non-hydrolyzable cap structure which can, for example, prevent decapping and increase RNA half-life. [0269] In some embodiments, a utilized 5' cap is a cap0, a cap1, or cap2 structure. See, e.g., Fig. 1 of Ramanathan A et al., and Fig. 1 of Decroly E et al., each of which is incorporated herein by reference in its entirety. In some embodiments, an RNA described herein comprises a cap1 structure. In some embodiments, an RNA described herein comprises a cap2 structure. [0270] In some embodiments, an RNA described herein comprises a cap0 structure. In some embodiments, a cap0 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G). In some embodiments, such a cap0 structure is connected to an RNA via a 5'- to 5'-triphosphate linkage and is also referred to herein as (m7)Gppp. In some embodiments, a cap0 structure comprises a guanosine nucleoside methylated at the 2'-position of the ribose of guanosine. In some embodiments, a cap0 structure comprises a guanosine nucleoside methylated at the 3'-position of the ribose of guanosine. In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine and at the 2'-position of the ribose ((m27,2'-O)G). In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine and at the 2'-position of the ribose ((m27,3'-O)G). [0271] In some embodiments, a cap1 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and optionally methylated at the 2' or 3' position of the ribose, and a 2'O methylated first nucleotide in an RNA ((m2'-O)N1). In some embodiments, a cap1 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and the 3' position of the ribose, and a 2'O methylated first nucleotide in an RNA ((m2'-O)N1). In some embodiments, a cap1 structure is connected to an RNA via a 5'- to 5'-triphosphate linkage and is 246 11979815v1 P-627574-PC also referred to herein as, e.g., ((m7)Gppp(2'-O)N1) or (m27,3’-O)Gppp(2'-O)N1), wherein N1 is as defined and described herein. In some embodiments, a cap1 structure comprises a second nucleotide, N2, which is at position 2 and is chosen from A, G, C, or U, e.g., (m7)Gppp(2'- O)N1pN2 or (m27,3’-O)Gppp(2'-O)N1pN2 , wherein each of N1 and N2 is as defined and described herein. [0272] In some embodiments, a cap2 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and optionally methylated at the 2' or 3' position of the ribose, and 2'O methylated first and second nucleotides in an RNA ((m2’-O)N1p(m2’-O)N2). In some embodiments, a cap2 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m7)G) and the 3' position of the ribose, and 2'O methylated first and second nucleotides in an RNA. In some embodiments, a cap2 structure is connected to an RNA via a 5'- to 5'- triphosphate linkage and is also referred to herein as, e.g., ((m7)Gppp(2'-O)N1p(2'-O)N2) or (m27,3’-O)Gppp(2'-O)N1p(2'-O)N2), wherein each of N1 and N2 is as defined and described herein. [0273] In some embodiments, the 5' cap is a dinucleotide cap structure. In some embodiments, the 5' cap is a dinucleotide cap structure comprising N1, wherein N1 is as defined and described herein. In some embodiments, the 5' cap is a dinucleotide cap G*N1, wherein N1 is as defined above and herein, and G* comprises a structure of formula (I):
Figure imgf000249_0001
wherein each R2 and R3 is -OH or -OCH3; and X is O or S. [0274] In some embodiments, R2 is -OH. In some embodiments, R2 is -OCH3. In some embodiments, R3 is -OH. In some embodiments, R3 is -OCH3. In some embodiments, R2 is -OH and R3 is -OH. In some embodiments, R2 is -OH and R3 is -CH3. In some embodiments, R2 is - CH3 and R3 is -OH. In some embodiments, R2 is -CH3 and R3 is -CH3. [0275] In some embodiments, X is O. In some embodiments, X is S. 247 11979815v1 P-627574-PC [0276] In some embodiments, the 5' cap is a dinucleotide cap0 structure (e.g., (m7)GpppN1, (m27,2’-O)GpppN1, (m27,3’-O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’- O)GppSpN1), wherein N1 is as defined and described herein. In some embodiments, the 5' cap is a dinucleotide cap0 structure (e.g., (m7)GpppN1, (m27,2’-O)GpppN1, (m27,3’-O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’-O)GppSpN1), wherein N1 is G. In some embodiments, the 5' cap is a dinucleotide cap0 structure (e.g., (m7)GpppN1, (m27,2’-O)GpppN1, (m27,3’-O)GpppN1, (m7)GppSpN1, (m27,2’-O)GppSpN1, or (m27,3’-O)GppSpN1), wherein N1 is A, U, or C. In some embodiments, the 5' cap is a dinucleotide cap1 structure (e.g., (m7)Gppp(m2’-O)N1, (m27,2’-O)Gppp(m2’-O)N1, (m27,3’-O)Gppp(m2’-O)N1, (m7)GppSp(m2’-O)N1, (m27,2’-O)GppSp(m2’-O)N1, or (m27,3’-O)GppSp(m2’-O)N1), wherein N1 is as defined and described herein. In some embodiments, the 5' cap is selected from the group consisting of (m7)GpppG (“Ecap0”), (m7)Gppp(m2’-O)G (“Ecap1”), (m2 7,3’-O)GpppG (“ARCA” or “D1”), and (m2 7,2’-O)GppSpG (“beta-S-ARCA”). In some embodiments, the 5' cap is (m7)GpppG (“Ecap0”), having a structure of formula (II):
Figure imgf000250_0001
[0277] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)G (“Ecap1”), having a structure of formula (III):
Figure imgf000250_0002
P-627574-PC [0278] In some embodiments, the 5' cap is (m2 7,3’-O)GpppG (“ARCA” or “D1”), having a structure of formula (IV): OH O O
Figure imgf000251_0001
some cap ARCA”), having a structure of formula (V): O OH O
Figure imgf000251_0002
[0280] In some embodiments, the 5' cap is a trinucleotide cap structure. In some embodiments, the 5' cap is a trinucleotide cap structure comprising N1pN2, wherein N1 and N2 are as defined and described herein. In some embodiments, the 5' cap is a dinucleotide cap G*N1pN2, wherein N1 and N2 are as defined above and herein, and G* comprises a structure of formula (VI):
Figure imgf000251_0003
defined and described herein. 249 11979815v1 P-627574-PC [0281] In some embodiments, the 5' cap is a trinucleotide cap0 structure (e.g., (m7)GpppN1pN2, (m2 7,2’-O)GpppN1pN2, or (m2 7,3’-O)GpppN1pN2), wherein N1 and N2 are as defined and described herein). In some embodiments, the 5' cap is a trinucleotide cap1 structure (e.g., (m7)Gppp(m2’- O)N1pN2, (m2 7,2’-O)Gppp(m2’-O)N1pN2, (m2 7,3’-O)Gppp(m2’-O)N1pN2), wherein N1 and N2 are as defined and described herein. In some embodiments, the 5' cap is a trinucleotide cap2 structure (e.g., (m7)Gppp(m2’-O)N1p(m2’-O)N2, (m2 7,2’-O)Gppp(m2’-O)N1p(m2’-O)N2, (m2 7,3’-O)Gppp(m2’- O)N1p(m2’-O)N2), wherein N1 and N2 are as defined and described herein. In some embodiments, the 5' cap is selected from the group consisting of (m27,3’-O)Gppp(m2’-O)ApG (“CleanCap AG”, “CC413”), (m2 7,3’-O)Gppp(m2’-O)GpG (“CleanCap GG”), (m7)Gppp(m2’-O)ApG, (m7)Gppp(m2’- O)GpG, (m2 7,3’-O)Gppp(m2 6,2’-O)ApG, and (m7)Gppp(m2’-O)ApU. [0282] In some embodiments, the 5' cap is (m2 7,3’-O)Gppp(m2’-O)ApG (“CleanCap AG”, “CC413”), having a structure of formula (VII):
Figure imgf000252_0001
[0283] In some embodiments, the 5' cap is (m2 7,3’-O)Gppp(m2’-O)GpG (“CleanCap GG”), having a structure of formula (VIII): 250 11979815v1 P-627574-PC
Figure imgf000253_0001
some cap a structure of formula (IX): OH OH NH2
Figure imgf000253_0002
[0285] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)GpG, having a structure of formula (X): 251 11979815v1 P-627574-PC
Figure imgf000254_0001
or a [0286] In some embodiments, the 5' cap is (m27,3’-O)Gppp(m26,2’-O)ApG, having a structure of formula (XI):
Figure imgf000254_0002
[0287] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)ApU, having a structure of formula (XII): 252 11979815v1 P-627574-PC
Figure imgf000255_0001
or a [0288] In some embodiments, the 5' cap is a tetranucleotide cap structure. In some embodiments, the 5' cap is a tetranucleotide cap structure comprising N1pN2pN3, wherein N1, N2, and N3 are as defined and described herein. In some embodiments, the 5' cap is a tetranucleotide cap G*N1pN2pN3, wherein N1, N2, and N3 are as defined above and herein, and G* comprises a structure of formula (XIII):
Figure imgf000255_0002
or a salt thereof, wherein R2, R3, and X are as defined and described herein. [0289] In some embodiments, the 5' cap is a tetranucleotide cap0 structure (e.g. (m7)GpppN1pN2pN3, (m2 7,2’-O)GpppN1pN2pN3, or (m2 7,3’-O)GpppN1N2pN3), wherein N1, N2, and N3 are as defined and described herein). In some embodiments, the 5’ cap is a tetranucleotide Cap1 structure (e.g., (m7)Gppp(m2’-O)N1pN2pN3, (m27,2’-O)Gppp(m2’-O)N1pN2pN3, (m27,3’-O)Gppp(m2’- O)N1pN2N3), wherein N1, N2, and N3 are as defined and described herein. In some embodiments, the 5' cap is a tetranucleotide Cap2 structure (e.g., (m7)Gppp(m2’-O)N1p(m2’-O)N2pN3, (m27,2’- O)Gppp(m2’-O)N1p(m2’-O)N2pN3, (m2 7,3’-O)Gppp(m2’-O)N1p(m2’-O)N2pN3), wherein N1, N2, and N3 are as defined and described herein. In some embodiments, the 5' cap is selected from the group 253 11979815v1 P-627574-PC consisting of (m2 7,3’-O)Gppp(m2’-O)Ap(m2’-O)GpG, (m2 7,3’-O)Gppp(m2’-O)Gp(m2’-O)GpC, (m7)Gppp(m2’-O)Ap(m2’-O)UpA, and (m7)Gppp(m2’-O)Ap(m2’-O)GpG. [0290] In some embodiments, the 5' cap is (m2 7,3’-O)Gppp(m2’-O)Ap(m2’-O)GpG, having a structure of formula (XIV):
Figure imgf000256_0001
[0291] In some embodiments, the 5' cap is (m27,3’-O)Gppp(m2’-O)Gp(m2’-O)GpC, having a structure of formula (XV): 254 11979815v1 P-627574-PC
Figure imgf000257_0001
some cap having a structure of formula (XVI):
Figure imgf000257_0002
P-627574-PC [0293] In some embodiments, the 5' cap is (m7)Gppp(m2’-O)Ap(m2’-O)GpG, having a structure of formula (XVII):
Figure imgf000258_0001
or a salt thereof. 2. Cap Proximal Sequences [0294] In some embodiments, a 5' UTR utilized in accordance with the present disclosure comprises a cap proximal sequence, e.g., as disclosed herein. In some embodiments, a cap proximal sequence comprises a sequence adjacent to a 5' cap. In some embodiments, a cap proximal sequence comprises nucleotides in positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide. [0295] In some embodiments, a cap structure comprises one or more polynucleotides of a cap proximal sequence. In some embodiments, a cap structure comprises an m7 guanosine cap and nucleotide +1 (N1) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m7 guanosine cap and nucleotide +2 (N2) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m7 guanosine cap and nucleotides +1 and +2 (N1 and N2) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m7 guanosine cap and nucleotides +1, +2, and +3 (N1, N2, and N3) of an RNA polynucleotide. [0296] Those skilled in the art, reading the present disclosure, will appreciate that, in some embodiments, one or more residues of a cap proximal sequence (e.g., one or more of residues +1, 256 11979815v1 P-627574-PC +2, +3, +4, and/or +5) may be included in an RNA by virtue of having been included in a cap entity (e.g., a cap1 or cap2 structure, etc.); alternatively, in some embodiments, at least some of the residues in a cap proximal sequence may be enzymatically added (e.g., by a polymerase such as a T7 polymerase). For example, in certain exemplified embodiments where a m27,3’- OGppp(m12’-O)ApG cap is utilized, +1 (i.e., N1) and +2 (i.e. N2) are the (m12’-O)A and G residues of the cap, and +3, +4, and +5 are added by a polymerase (e.g., T7 polymerase). [0297] In some embodiments, the 5'’ cap is a dinucleotide cap structure, wherein the cap proximal sequence comprises N1 of the 5’ cap, where N1 is any nucleotide, e.g., A, C, G or U. In some embodiments, the 5' cap is a trinucleotide cap structure (e.g., the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N1 and N2 of the 5' cap, wherein N1 and N2 are independently any nucleotide, e.g., A, C, G or U. In some embodiments, the 5' cap is a tetranucleotide cap structure (e.g., the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N1, N2, and N3 of the 5' cap, wherein N1, N2, and N3 are any nucleotide, e.g., A, C, G or U. [0298] In some embodiments, e.g., where the 5' cap is a dinucleotide cap structure, a cap proximal sequence comprises N1 of a the 5' cap, and N2, N3, N4 and N5, wherein N1 to N5 correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide. In some embodiments, e.g., where the 5' cap is a trinucleotide cap structure, a cap proximal sequence comprises N1 and N2 of a the 5' cap, and N3, N4 and N5, wherein N1 to N5 correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide. In some embodiments, e.g., where the 5' cap is a tetranucleotide cap structure, a cap proximal sequence comprises N1, N2, and N3 of a the 5’ cap, and N4 and N5, wherein N1 to N5 correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide. [0299] In some embodiments, N1 is A. In some embodiments, N1 is C. In some embodiments, N1 is G. In some embodiments, N1 is U. In some embodiments, N2 is A. In some embodiments, N2 is C. In some embodiments, N2 is G. In some embodiments, N2 is U. In some embodiments, N3 is A. In some embodiments, N3 is C. In some embodiments, N3 is G. In some embodiments, N3 is U. In some embodiments, N4 is A. In some embodiments, N4 is C. In some embodiments, N4 is G. In some embodiments, N4 is U. In some embodiments, N5 is A. In some embodiments, N5 is C. In some embodiments, N5 is G. In some embodiments, N5 is U. It will be understood that, each of the embodiments described above and herein (e.g., for N1 through N5) may be taken singly or in combination and/or may be combined with other embodiments of variables described above and herein (e.g., 5' caps). 257 11979815v1 P-627574-PC [0300] In some embodiments, a cap proximal sequence comprises A1 and G2 of the Cap1 structure, and a sequence comprising: A3A4U5 at positions +3, +4 and +5 respectively of the nucleotide sequence. 3. 5’ UTR [0301] In some embodiments, an RNA utilized in accordance with the present disclosure comprises a 5'-UTR. In some embodiments, a 5’-UTR may comprise a plurality of distinct sequence elements; in some embodiments, such plurality may be or comprise multiple copies of one or more particular sequence elements (e.g., as may be from a particular source or otherwise known as a functional or characteristic sequence element). In some embodiments, a 5’ UTR comprises multiple different sequence elements. [0302] The term “untranslated region” or “UTR” is commonly used in the art to refer to a region in a DNA molecule which is transcribed but is not translated into an amino acid sequence, or to the corresponding region in an RNA polynucleotide, such as an mRNA molecule. An untranslated region (UTR) can be present 5' (upstream) of an open reading frame (5'-UTR) and/or 3' (downstream) of an open reading frame (3'-UTR). As used herein, the terms “five prime untranslated region” or “5' UTR” refer to a sequence of a nucleotide sequence between the 5' end of the nucleotide sequence (e.g., a transcription start site) and a start codon of a coding region of the nucleotide sequence. In some embodiments, “5' UTR” refers to a sequence of a nucleotide sequence that begins at the 5' end of the nucleotide sequence (e.g., a transcription start site) and ends one nucleotide (nt) before a start codon (usually AUG) of a coding region of the nucleotide sequence, e.g., in its natural context. In some embodiments, a 5' UTR comprises a Kozak sequence. A 5'-UTR is downstream of the 5'-cap (if present), e.g., directly adjacent to the 5'-cap. In some embodiments, a 5' UTR disclosed herein comprises a cap proximal sequence, e.g., as defined and described herein. In some embodiments, a cap proximal sequence comprises a sequence adjacent to a 5' cap. [0303] Exemplary 5' UTRs include a human alpha globin (hAg) 5'UTR or a fragment thereof, a TEV 5' UTR or a fragment thereof, a HSP705' UTR or a fragment thereof, or a c-Jun 5' UTR or a fragment thereof. [0304] In some embodiments, an RNA disclosed herein comprises a hAg 5' UTR or a fragment thereof. [0305] In some embodiments, an RNA disclosed herein comprises a 5' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% 258 11979815v1 P-627574-PC identity to a 5' UTR with the sequence AGAATAAACTAGTATTCTTCTGGTCCCCACAGACTCAGAGAGAACCCGCCACC (SEQ ID NO: 210). In some embodiments, an RNA disclosed herein comprises a 5' UTR having the sequence as set forth in SEQ ID NO: 210. [0306] In some embodiments, an RNA disclosed herein comprises a 5' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 5’ UTR with the sequence AACUAGUAUUCUUCUGGUCCCCACAGACUCAGAGAGAACCCGCCACC (SEQ ID NO: 211) (hAg-Kozak/5'UTR). In some embodiments, an RNA disclosed herein comprises a 5' UTR having the sequence as set forth in SEQ ID NO 211. 4. PolyA Tail [0307] In some embodiments, a polynucleotide (e.g., DNA, RNA) disclosed herein comprises a polyadenylate (polyA) sequence, e.g., as described herein. In some embodiments, a polyA sequence is situated downstream of a 3'-UTR, e.g., adjacent to a 3'-UTR. [0308] As used herein, the term “poly(A) sequence” or “poly-A tail” refers to an uninterrupted or interrupted sequence of adenylate residues which is typically located at the 3'-end of an RNA polynucleotide. Poly(A) sequences are known to those of skill in the art and may follow the 3’- UTR in the RNAs described herein. An uninterrupted poly(A) sequence is characterized by consecutive adenylate residues. In nature, an uninterrupted poly(A) sequence is typical. In some embodiments, polynucleotides disclosed herein comprise an uninterrupted poly(A) sequence. In some embodiments, polynucleotides disclosed herein comprise interrupted poly(A) sequence. In some embodiments, RNAs disclosed herein can have a poly(A) sequence attached to the free 3'- end of the RNA by a template-independent RNA polymerase after transcription or a poly(A) sequence encoded by DNA and transcribed by a template-dependent RNA polymerase. [0309] It has been demonstrated that a poly(A) sequence of about 120 A nucleotides has a beneficial influence on the levels of RNA in transfected eukaryotic cells, as well as on the levels of protein that are translated from an open reading frame that is present upstream (5') of the poly(A) sequence (Holtkamp et al., 2006, Blood, vol.108, pp.4009-4017, which is herein incorporated by reference). [0310] In some embodiments, a poly(A) sequence in accordance with the present disclosure is not limited to a particular length; in some embodiments, a poly(A) sequence is any length. In some embodiments, a poly(A) sequence comprises, essentially consists of, or consists of at least 20, at 259 11979815v1 P-627574-PC least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 A nucleotides, and, in particular, about 120 A nucleotides. In this context, "essentially consists of" means that most nucleotides in the poly(A) sequence, typically at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by number of nucleotides in the poly(A) sequence are A nucleotides, but permits that remaining nucleotides are nucleotides other than A nucleotides, such as U nucleotides (uridylate), G nucleotides (guanylate), or C nucleotides (cytidylate). In this context, "consists of" means that all nucleotides in the poly(A) sequence, i.e., 100% by number of nucleotides in the poly(A) sequence, are A nucleotides. The term “A nucleotide” or “A” refers to adenylate. [0311] In some embodiments, a poly(A) sequence is attached during RNA transcription, e.g., during preparation of in vitro transcribed RNA, based on a DNA template comprising repeated dT nucleotides (deoxythymidylate) in the strand complementary to the coding strand. The DNA sequence encoding a poly(A) sequence (coding strand) is referred to as a poly(A) cassette. [0312] In some embodiments, the poly(A) cassette present in the coding strand of DNA essentially consists of dA nucleotides, but is interrupted by a random sequence of the four nucleotides (dA, dC, dG, and dT). Such a random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides in length. Such a cassette is disclosed in WO 2016/005324 A1, hereby incorporated by reference. Any poly(A) cassette disclosed in WO 2016/005324 A1, which is incorporated herein by reference in its entirety, may be used in accordance with the present disclosure. A poly(A) cassette that essentially consists of dA nucleotides, but is interrupted by a random sequence having an equal distribution of the four nucleotides (dA, dC, dG, dT) and having a length of e.g., 5 to 50 nucleotides shows, at the DNA level, constant propagation of plasmid DNA in E. coli and is still associated, at the RNA level, with the beneficial properties with respect to supporting RNA stability and translational efficiency is encompassed. In some embodiments, the poly(A) sequence contained in an RNA polynucleotide described herein essentially consists of A nucleotides, but is interrupted by a random sequence of the four nucleotides (A, C, G, U). Such a random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides in length. [0313] In some embodiments, no nucleotides other than A nucleotides flank a poly(A) sequence at its 3'-end, i.e., the poly(A) sequence is not masked or followed at its 3'-end by a nucleotide other than A. [0314] In some embodiments, the poly(A) sequence may comprise at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 260 11979815v1 P-627574-PC nucleotides. In some embodiments, the poly(A) sequence may essentially consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence may consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence comprises at least 100 nucleotides. In some embodiments, the poly(A) sequence comprises about 150 nucleotides. In some embodiments, the poly(A) sequence comprises about 120 nucleotides. [0315] In some embodiments, a poly(A) sequence comprises a specific number of adenosines, such as about 50 or more, about 60 or more, about 70 or more, about 80 or more, about 90 or more, about 100 or more, about 120, or about 150 or about 200. In some embodiments a poly(A) sequence of an RNA may comprise 200 A residues or less. In some embodiments, a poly(A) sequence of an RNA may comprise about 200 A residues. In some embodiments, a poly(A) sequence of an RNA may comprise 180 A residues or less. In some embodiments, a poly(A) sequence of an RNA may comprise about 180 A residues. In some embodiments, a poly(A) sequence may comprise 150 residues or less. [0316] In some embodiments, RNA comprises a poly(A) sequence comprising the nucleotide sequence of: AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGCATATGACTAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAA (SEQ ID NO: 212), or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of: SEQ ID NO: 212. In some embodiments, a poly(A) sequence comprises a plurality of A residues interrupted by a linker. In some embodiments, a linker comprises the nucleotide sequence GCATATGAC. [0317] In some embodiments, an RNA comprises a poly(A) sequence comprising the nucleotide sequence of: AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGCAUAUGACUAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAA (SEQ ID NO: 214), or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 214. In some embodiments, a poly(A) sequence comprises a plurality of A residues interrupted by a linker. In some embodiments, a linker comprises the nucleotide sequence GCAUAUGAC. 261 11979815v1 P-627574-PC 5. 3' UTR [0318] In some embodiments, an RNA utilized in accordance with the present disclosure comprises a 3'-UTR. As used herein, the terms “three prime untranslated region,” “3' untranslated region,” or “3' UTR” refer to a sequence of an mRNA molecule that begins following a stop codon of a coding region of an open reading frame sequence. In some embodiments, the 3' UTR begins immediately after a stop codon of a coding region of an open reading frame sequence, e.g., in its natural context. In other embodiments, the 3' UTR does not begin immediately after stop codon of the coding region of an open reading frame sequence, e.g., in its natural context. The term “3'- UTR” preferably does not include the poly(A) sequence. Thus, the 3'-UTR is upstream of the poly(A) sequence (if present), e.g. directly adjacent to the poly(A) sequence. [0319] In some embodiments, an RNA disclosed herein comprises a 3'UTR comprising an F element and/or an I element. In some embodiments, a 3' UTR or a proximal sequence thereto comprises a restriction site. In some embodiments, a restriction site is a BamHI site. In some embodiments, a restriction site is an XhoI site. [0320] In some embodiments, an RNA construct comprises an F element. In some embodiments, an F element sequence is a 3' UTR of amino-terminal enhancer of split (AES). [0321] In some embodiments, an RNA disclosed herein comprises a 3' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 3’ UTR with the sequence of CTGGTACTGCATGCACGCAATGCTAGCTGCCCCTTTCCCGTCCTGGGTACCCCGAGT CTCCCCCGACCTCGGGTCCCAGGTATGCTCCCACCTCCACCTGCCCCACTCACCACC TCTGCTAGTTCCAGACACCTCCCAAGCACGCAGCAATGCAGCTCAAAACGCTTAGCC TAGCCACACCCCCACGGGAAACAGCAGTGATTAACCTTTAGCAATAAACGAAAGTT TAACTAAGCTATACTAACCCCAGGGTTGGTCAATTTCGTGCCAGCCACACC (SEQ ID NO: 216). In some embodiments, an RNA disclosed herein comprises a 3' UTR with the sequence of SEQ ID NO: 216. [0322] In some embodiments, an RNA disclosed herein comprises a 3' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 3’ UTR with the sequence of CUGGUACUGCAUGCACGCAAUGCUAGCUGCCCCUUUCCCGUCCUGGGUACCCCGA GUCUCCCCCGACCUCGGGUCCCAGGUAUGCUCCCACCUCCACCUGCCCCACUCACC ACCUCUGCUAGUUCCAGACACCUCCCAAGCACGCAGCAAUGCAGCUCAAAACGCU 262 11979815v1 P-627574-PC UAGCCUAGCCACACCCCCACGGGAAACAGCAGUGAUUAACCUUUAGCAAUAAACG AAAGUUUAACUAAGCUAUACUAACCCCAGGGUUGGUCAAUUUCGUGCCAGCCACA CC (SEQ ID NO: 217). In some embodiments, an RNA disclosed herein comprises a 3' UTR with the sequence of SEQ ID NO: 217. [0323] In some embodiments, a 3' UTR is an FI element as described in WO2017/060314, which is herein incorporated by reference in its entirety. B. Modified RNAs [0324] In some embodiments, the present disclosure provides compositions comprising modified RNAs and methods of use thereof. In some embodiments, the modified RNA comprises one or more modified nucleoside residues. For example, in some embodiments, an RNA comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 2, Table 4, or Table 6, comprises one or more modified nucleoside residues. [0325] In some embodiments, an RNA as described herein refers to a messenger RNA. [0326] In some embodiments, all uridine residues are modified as described herein. In some embodiments, one or more of the RNAs as described herein are nucleoside modified RNAs. In other embodiments, two or more of the RNAs as described herein are nucleoside modified RNAs. In other embodiments, three or more of the RNAs as described herein are nucleoside modified RNAs. [0327] In some embodiments, the modified nucleoside of the methods and compositions of the present disclosure is m5C (5-methylcytidine). In another embodiment, the modified nucleoside is m5U (5-methyluridine). In another embodiment, the modified nucleoside is m6A (N6- methyladenosine). In another embodiment, the modified nucleoside is s2U (2-thiouridine). In another embodiment, the modified nucleoside is Ψ (pseudouridine). In another embodiment, the modified nucleoside is Um (2'-O-methyluridine). [0328] In some embodiments, a modified nucleoside is m1A (1-methyladenosine), m2A (2- methyladenosine), m6A (N6-methyladenosine), Am (2'-O-methyladenosine), ms2m6A (2- methylthio-N6-methyladenosine), i6A (N6-isopentenyladenosine), ms2i6A (2-methylthio-N6- isopentenyladenosine), io6A (N6-(cis-hydroxyisopentenyl)adenosine), ms2io6A (2-methylthio- N6-(cis-hydroxyisopentenyl) adenosine), g6A (N6-glycinylcarbamoyladenosine), t6A (N6- threonylcarbamoyladenosine), ms2t6A (2-methylthio-N6-threonyl carbamoyladenosine), m6t6A 263 11979815v1 P-627574-PC (N6-methyl-N6-threonylcarbamoyladenosine), hn6A (N6-hydroxynorvalylcarbamoyladenosine), ms2hn6A (2-methylthio-N6-hydroxynorvalyl carbamoyladenosine), Ar(p) (2'-O-ribosyladenosine (phosphate)), I (inosine), m1I (1-methylinosine), m1Im (1,2'-O-dimethylinosine), m3C (3- methylcytidine), m5C (5-methylcytidine), Cm (2'-O-methylcytidine), s2C (2-thiocytidine), ac4C (N4-acetylcytidine), f5C (5-formylcytidine), m5Cm (5,2'-O-dimethylcytidine), ac4Cm (N4-acetyl- 2'-O-methylcytidine), k2C (lysidine), m1G (1-methylguanosine), m2G (N2-methylguanosine), m7G (7-methylguanosine), Gm (2'-O-methylguanosine), m2 2G (N2,N2-dimethylguanosine), m2Gm (N2,2'-O-dimethylguanosine), m22Gm (N2,N2,2'-O-trimethylguanosine), Gr(p) (2'-O- ribosylguanosine (phosphate)), yW (wybutosine), o2yW (peroxywybutosine), OHyW (hydroxywybutosine), OHyW* (undermodified hydroxywybutosine), imG (wyosine), mimG (methylwyosine), Q (queuosine), oQ (epoxyqueuosine), galQ (galactosyl-queuosine), manQ (mannosyl-queuosine), preQ0 (7-cyano-7-deazaguanosine), preQ1 (7-aminomethyl-7- deazaguanosine), G+ (archaeosine), ^ (pseudouridine), D (dihydrouridine), m5U (5- methyluridine), Um (2'-O-methyluridine), m5Um (5,2'-O-dimethyluridine), m1 ^ (1- methylpseudouridine), ^m (2'-O-methylpseudouridine), s2U (2-thiouridine), s4U (4-thiouridine), m5s2U (5-methyl-2-thiouridine), s2Um (2-thio-2'-O-methyluridine), acp3U (3-(3-amino-3- carboxypropyl)uridine), ho5U (5-hydroxyuridine), mo5U (5-methoxyuridine), cmo5U (uridine 5- oxyacetic acid), mcmo5U (uridine 5-oxyacetic acid methyl ester), chm5U (5- (carboxyhydroxymethyl)uridine), mchm5U (5-(carboxyhydroxymethyl)uridine methyl ester), mcm5U (5-methoxycarbonylmethyluridine), mcm5Um (5-methoxycarbonylmethyl-2'-O- methyluridine), mcm5s2U (5-methoxycarbonylmethyl-2-thiouridine), nm5s2U (5-aminomethyl-2- thiouridine), mnm5U (5-methylaminomethyluridine), mnm5s2U (5-methylaminomethyl-2- thiouridine), mnm5se2U (5-methylaminomethyl-2-selenouridine), ncm5U (5- carbamoylmethyluridine), ncm5Um (5-carbamoylmethyl-2'-O-methyluridine), cmnm5U (5- carboxymethylaminomethyluridine), cmnm5Um (5-carboxymethylaminomethyl- 2'-O- methyluridine), cmnm5s2U (5-carboxymethylaminomethyl-2-thiouridine), m62A (N6,N6- dimethyladenosine), Im (2'-O-methylinosine), m4C (N4-methylcytidine), m4Cm (N4,2'-O- dimethylcytidine), hm5C (5-hydroxymethylcytidine), m3U (3-methyluridine), m1acp3 ^ (1-methyl- 3-(3-amino-3-carboxypropyl) pseudouridine), cm5U (5-carboxymethyluridine), m6Am (N6,2'-O- dimethyladenosine), m6 2Am (N6,N6,2'-O-trimethyladenosine), m2,7G (N2,7-dimethylguanosine), m2,2,7G (N2,N2,7-trimethylguanosine), m3Um (3,2'-O-dimethyluridine), m5D (5- methyldihydrouridine), m3 ^ (3-methylpseudouridine), f5Cm (5-formyl-2'-O-methylcytidine), 264 11979815v1 P-627574-PC m1Gm (1,2'-O-dimethylguanosine), m1Am (1,2'-O-dimethyladenosine), ^m5U (5- taurinomethyluridine), ^m5s2U (5-taurinomethyl-2-thiouridine), imG-14 (4-demethylwyosine), imG2 (isowyosine), ac6A (N6-acetyladenosine), inm5U (5-(isopentenylaminomethyl)uridine), inm5s2U (5-(isopentenylaminomethyl)- 2-thiouridine), inm5Um (5-(isopentenylaminomethyl)- 2'- O-methyluridine), m2,7Gm (N2,7,2'-O-trimethylguanosine), m4 2Cm (N4,N4,2'-O- trimethylcytidine), C+ (agmatidine), m8A (8-methyladenosine), gmnm5s2U (geranylated 5- methylaminomethyl-2-thiouridine), gcmnm5s2U (geranylated 5-carboxymethylaminomethyl-2- thiouridine), or cnm5U (5-cyanomethyl-uridine). [0329] In some embodiments, modified nucleoside residues are pseudouridine or pseudouridine family residues. [0330] In some embodiments, the modified RNA comprises pseudouridine residues. In some embodiments, pseudouridine refers to the C-glycoside isomer of the nucleoside uridine. In some embodiments, pseudouridine residues comprise m1acp3Ψ (1-methyl-3-(3-amino-5- carboxypropyl)pseudouridine, m1Ψ (1-methylpseudouridine), Ψm (2′-O-methylpseudouridine, m5D (5-methyldihydrouridine), m3Ψ (3-methylpseudouridine), or a combination thereof. In some embodiments, said pseudouridine residues comprise 1-methylpseudouridine residues instead of uridine. [0331] In some embodiments, modified nucleoside residues are pseudouridine analogues. In some embodiments, a "pseudouridine analog" is any modification, variant, isoform or derivative of pseudouridine. For example, pseudouridine analogs include but are not limited to 1- carboxymethyl-pseudouridine, 1-propynyl-pseudouridine, 1-taurinomethyl-pseudouridine, 1- taurinomethyl-4-thio-pseudouridine, 1-methylpseudouridine (m1 ^), 1-methyl-4-thio- pseudouridine (m1s4 ^), 4-thio-1-methyl-pseudouridine, 3-methyl-pseudouridine (m3 ^), 2-thio-1- methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydropseudouridine, 2-thio-dihydropseudouridine, 2-methoxyuridine, 2-methoxy-4-thio- uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, N1-methyl-pseudouridine, 1-methyl-3-(3-amino-3-carboxypropyl)pseudouridine (acp3 ^), and 2'-O-methyl-pseudouridine ( ^m). [0332] In some embodiments, a modified nucleobase is a modified uracil. Exemplary nucleobases and nucleosides having a modified uracil include pseudouridine ( ^), pyridin-4-one ribonucleoside, 5-aza-uridine, 6-aza-uridine, 2-thio-5-aza-uridine, 2-thio-uridine (s2U), 4-thio- uridine (s4U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxy-uridine (ho5U), 5- 265 11979815v1 P-627574-PC aminoallyl-uridine, 5-halo-uridine (e.g., 5-iodo-uridine or 5-bromo-uridine), 3-methyl-uridine (m3U), 5-methoxy-uridine (mo5U), uridine 5-oxyacetic acid (cmo5U), uridine 5-oxyacetic acid methyl ester (mcmo5U), 5-carboxymethyl-uridine (cm5U), 1-carboxymethyl-pseudouridine, 5- carboxyhydroxymethyl-uridine (chm5U), 5-carboxyhydroxymethyl-uridine methyl ester (mchm5U), 5-methoxycarbonylmethyl-uridine (mcm5U), 5-methoxycarbonylmethyl-2-thio- uridine (mcm5s2U), 5-aminomethyl-2-thio-uridine (nm5s2U), 5-methylaminomethyl-uridine (mnm5U), 5-methylaminomethyl-2-thio-uridine (mnm5s2U), 5-methylaminomethyl-2-seleno- uridine (mnm5se2U), 5-carbamoylmethyl-uridine (ncm5U), 5-carboxymethylaminomethyl-uridine (cmnm5U), 5-carboxymethylaminomethyl-2-thio-uridine (cmnm5s2U), 5-propynyl-uridine, 1- propynyl-pseudouridine, 5-taurinomethyl-uridine ( ^cm5U), 1-taurinomethyl-pseudouridine, 5- taurinomethyl-2-thio-uridine ( ^rm5s2U), 1-taurinomethyl-4-thio-pseudouridine, 5-methyl-uridine (m5U, i.e., having the nucleobase deoxythymine), 1-methylpseudouridine (m1 ^), 5-methyl-2-thio- uridine (m5s2U), 1-methyl-4-thio-pseudouridine (m1s4 ^), 4-thio-1-methyl-pseudouridine, 3- methyl-pseudouridine (m3 ^), 2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine (D), dihydropseudouridine, 5,6- dihydrouridine, 5-methyl-dihydrouridine (m5D), 2-thio-dihydrouridine, 2-thio- dihydropseudouridine, 2-methoxy-uridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, N1-methyl-pseudouridine (also known as 1- methylpseudouridine (m1 ^), 3-(3-amino-3-carboxypropyl)uridine (acp3U), 1-methyl-3-(3-amino- 3-carboxypropyl)pseudouridine (acp 3 ^), 5-(isopentenylaminomethyl)uridine (inm 5 U), 5- (isopentenylaminomethyl)-2-thio-uridine (inm5s2U), α-thio-uridine, 2'-O-methyl-uridine (Um), 5,2'-O-dimethyl-uridine (m5Um), 2'-O-methyl-pseudouridine ( ^m), 2-thio-2'-O-methyl-uridine (s2Um), 5-methoxycarbonylmethyl-2'-O-methyl-uridine (mcm5Um), 5-carbamoylmethyl-2'-β- methyl-uridine (ncm5Um), 5-carboxymethylaminomethyl-2'-O-methyl-uridine (cmnm5Um), 3,2'- O-dimethyl-uridine (m 3 Um), 5-(isopentenylaminomethyl)-2'-β-methyl-uridine (inm 5 Um), 1-thio- uridine, deoxythymidine, 2'-F-ara-uridine, 2'-F-uridine, 2'-OH-ara-uridine, 5-(2- carbomethoxyvinyl)uridine, and 5-[3-(1-E-propenylamino)uridine. [0333] In some embodiments, a modified nucleobase is a modified cytosine. Exemplary nucleobases and nucleosides having a modified cytosine include 5-aza-cytidine, 6-aza-cytidine, pseudoisocytidine, 3-methyl-cytidine (m3C), N4-acetyl-cytidine (ac4C), 5-formyl-cytidine (f5C), N4-methyl-cytidine (m4C), 5-methyl-cytidine (m5C), 5-halo-cytidine (e.g., 5-iodo-cytidine), 5- hydroxymethyl-cytidine (hm5C), 1-methyl-pseudoisocytidine, pyrrolo-cytidine, pyrrolo- 266 11979815v1 P-627574-PC pseudoisocytidine, 2-thio-cytidine (s2C), 2-thio-5-methyl-cytidine, 4-thio-pseudoisocytidine, 4- thio-1-methyl-pseudoisocytidine, 4-thio-1-methyl-1-deaza-pseudoisocytidine, 1-methyl-1-deaza- pseudoisocytidine, zebularine, 5-aza-zebularine, 5-methyl-zebularine, 5-aza-2-thio-zebularine, 2- thio-zebularine, 2-methoxy-cytidine, 2-methoxy-5-methyl-cytidine, 4-methoxy- pseudoisocytidine, 4-methoxy-1-methyl-pseudoisocytidine, lysidine (k2C), α-thio-cytidine, 2'-O- methyl-cytidine (Cm), 5,2'-O-dimethyl-cytidine (m5Cm), N4-acetyl-2'-O-methyl-cytidine (ac4Cm), N4,2'-O-dimethyl-cytidine (m4Cm), 5-formyl-2'-O-methyl-cytidine (f5Cm), N4,N4,2'-O- trimethyl-cytidine (m42Cm), 1-thio-cytidine, 2'-F-ara-cytidine, 2'-F-cytidine, and 2'-OH-ara- cytidine. [0334] In some embodiments, a modified nucleobase is a modified adenine. Exemplary nucleobases and nucleosides having a modified adenine include 2-amino-purine, 2,6- diaminopurine, 2-amino-6-halo-purine (e.g., 2-amino-6-chloro-purine), 6-halo-purine (e.g., 6- chloro-purine), 2-amino-6-methyl-purine, 8-azido-adenosine, 7-deaza-adenine, 7-deaza-8-aza- adenine, 7-deaza-2-amino-purine, 7-deaza-8-aza-2-amino-purine, 7-deaza-2,6-diaminopurine, 7- deaza-8-aza-2,6-diaminopurine, 1-methyl-adenosine (m1A), 2-methyl-adenine (m2A), N6-methyl- adenosine (m6A), 2-methylthio-N6-methyl-adenosine (ms2 m6A), N6-isopentenyl-adenosine (i6A), 2-methylthio-N6-isopentenyl-adenosine (ms2i6A), N6-(cis-hydroxyisopentenyl)adenosine (io6A), 2-methylthio-N6-(cis-hydroxyisopentenyl)adenosine (ms2io6A), N6-glycinylcarbamoyl- adenosine (g6A), N6-threonylcarbamoyl-adenosine (t6A), N6-methyl-N6-threonylcarbamoyl- adenosine (m6t6A), 2-methylthio-N6-threonylcarbamoyl-adenosine (ms2g6A), N6,N6-dimethyl- adenosine (m6 2A), N6-hydroxynorvalylcarbamoyl-adenosine (hn6A), 2-methylthio-N6- hydroxynorvalylcarbamoyl-adenosine (ms2hn6A), N6-acetyl-adenosine (ac6A), 7-methyl-adenine, 2-methylthio-adenine, 2-methoxy-adenine, α-thio-adenosine, 2'-O-methyl-adenosine (Am), N6,2'- O-dimethyl-adenosine (m6Am), N6,N6,2'-O-trimethyl-adenosine (m6 2Am), 1,2'-O-dimethyl- adenosine (m1Am), 2'-β-ribosyladenosine (phosphate) (Ar(p)), 2-amino-N6-methyl-purine, 1- thio-adenosine, 8-azido-adenosine, 2'-F-ara-adenosine, 2'-F-adenosine, 2'-OH-ara-adenosine, and N6-(19-amino-pentaoxanonadecyl)-adenosine. [0335] In some embodiments, a modified nucleobase is a modified guanine. Exemplary nucleobases and nucleosides having a modified guanine include inosine (I), 1-methyl-inosine (m1I), wyosine (imG), methylwyosine (mimG), 4-demethyl-wyosine (imG-14), isowyosine (imG2), wybutosine (yW), peroxywybutosine (o2yW), hydroxywybutosine (OHyW), undermodified hydroxywybutosine (OHyW*), 7-deaza-guanosine, queuosine (Q), 267 11979815v1 P-627574-PC epoxyqueuosine (oQ), galactosyl-queuosine (galQ), mannosyl-queuosine (manQ), 7-cyano-7- deaza-guanosine (preQ0), 7-aminomethyl-7-deaza-guanosine (preQ1), archaeosine (G+), 7-deaza- 8-aza-guanosine, 6-thio-guanosine, 6-thio-7-deaza-guanosine, 6-thio-7-deaza-8-aza-guanosine, 7- methyl-guanosine (m7G), 6-thio-7-methyl-guanosine, 7-methyl-inosine, 6-methoxy-guanosine, 1- methyl-guanosine (m1G), N2-methyl-guanosine (m2G), N2,N2-dimethyl-guanosine (m2 2G), N2,7- dimethyl-guanosine (m2,7G), N2,N2,7-dimethyl-guanosine (m2,2,7G), 8-oxo-guanosine, 7- methyl-8-oxo-guanosine, 1-methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine, N2,N2- dimethyl-6-thio-guanosine, α-thio-guanosine, 2'-O-methyl-guanosine (Gm), N2-methyl-2'-O- methyl-guanosine (m2Gm), N2,N2-dimethyl-2'-O-methyl-guanosine (m2 2Gm), 1-methyl-2'-O- methyl-guanosine (m1Gm), N2,7-dimethyl-2'-O-methyl-guanosine (m2'7Gm), 2'-O-methyl-inosine (Im), 1,2'-O-dimethyl-inosine (m1Im), and 2'-O-ribosylguanosine (phosphate) (Gr(p)). [0336] The nucleobase of the nucleotide can be independently selected from a purine, a pyrimidine, a purine or pyrimidine analog. For example, the nucleobase can each be independently selected from adenine, cytosine, guanine, uracil, or hypoxanthine. In another embodiment, the nucleobase can also include, for example, naturally-occurring and synthetic derivatives of a base, including pyrazolo[3,4-d]pyrimidines, 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo (e.g., 8-bromo), 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, deazaguanine, 7-deazaguanine, 3-deazaguanine, deazaadenine, 7-deazaadenine, 3- deazaadenine, pyrazolo[3,4-d]pyrimidine, imidazo[1,5-a]1,3,5 triazinones, 9-deazapurines, imidazo[4,5-d]pyrazines, thiazolo[4,5-d]pyrimidines, pyrazin-2-ones, 1,2,4-triazine, pyridazine; and 1,3,5 triazine. When the nucleotides are depicted using the shorthand A, G, C, T or U, each letter refers to the representative base and/or derivatives thereof, e.g., A includes adenine or adenine analogs, e.g., 7-deaza adenine). Modifications on the Internucleoside Linkage [0337] The modified nucleotides, which may be incorporated into a polynucleotide, primary construct, or RNA molecule, can be modified on the internucleoside linkage (e.g., phosphate backbone). Herein, in the context of the polynucleotide backbone, the phrases "phosphate" and 268 11979815v1 P-627574-PC "phosphodiester" are used interchangeably. Backbone phosphate groups can be modified by replacing one or more of the oxygen atoms with a different substituent. Further, the modified nucleosides and nucleotides can include the wholesale replacement of an unmodified phosphate moiety with another internucleoside linkage as described herein. Examples of modified phosphate groups include, but are not limited to, phosphorothioate, phosphoroselenates, boranophosphates, boranophosphate esters, hydrogen phosphonates, phosphoramidates, phosphorodiamidates, alkyl or aryl phosphonates, and phosphotriesters. Phosphorodithioates have both non-linking oxygens replaced by sulfur. The phosphate linker can also be modified by the replacement of a linking oxygen with nitrogen (bridged phosphoramidates), sulfur (bridged phosphorothioates), and carbon (bridged methylene-phosphonates). [0338] The α-thio substituted phosphate moiety is provided to confer stability to RNA and DNA polymers through the unnatural phosphorothioate backbone linkages. Phosphorothioate DNA and RNA have increased nuclease resistance and subsequently a longer half-life in a cellular environment. Phosphorothioate linked polynucleotides, primary constructs, or modified RNA molecules are expected to also reduce the innate immune response through weaker binding/activation of cellular innate immune molecules. [0339] In specific embodiments, a modified nucleoside includes an alpha-thio-nucleoside (e.g., 5'- O-(1-thiophosphate)-adenosine, 5'-O-(1-thiophosphate)-cytidine (α-thio-cytidine), 5'-O-(1- thiophosphate)-guanosine, 5'-O-(1-thiophosphate)-uridine, or 5'-O-(1-thiophosphate)- pseudouridine). [0340] Other internucleoside linkages that may be employed according to the present disclosure, including internucleoside linkages which do not contain a phosphorous atom, are described herein below. Combinations of Modified Sugars, Nucleobases, and Internucleoside Linkages [0341] The polynucleotides, primary constructs, and modified RNA of the disclosure can include a combination of modifications to the sugar, the nucleobase, and/or the internucleoside linkage. [0342] In another embodiment, the purified preparation of RNA, oligoribonucleotide, or polyribonucleotide of the methods and compositions of the present disclosure comprises a combination of two or more of the above-described modifications. In another embodiment, the purified preparation of the RNA or oligoribonucleotide comprises a combination of three or more of the above-described modifications. In another embodiment, the purified preparation of the RNA 269 11979815v1 P-627574-PC or oligoribonucleotide comprises a combination of more than three of the above-described modifications. [0343] In some embodiments, the modified RNAs comprise in vitro-synthesized modified RNAs. [0344] In some embodiments, the present disclosure comprises one or more modified RNAs encoding an HSV glycoprotein. In some embodiments, the modified RNA comprises pseudouridine or pseudouridine family residues. In another embodiment, the modified RNAs of the present disclosure are capable of directing protein expression of HSV glycoproteins encoded thereon. [0345] In another embodiment, the present disclosure provides an in vitro-transcribed mRNA molecule encoding an HSV glycoprotein, comprising a pseudouridine. In another embodiment, the present disclosure provides a synthetic RNA molecule encoding an HSV glycoprotein, comprising a pseudouridine. [0346] In another embodiment, an in vitro-transcribed RNA molecule of the methods and compositions of the present disclosure is synthesized by T7 phage RNA polymerase. In another embodiment, the molecule is synthesized by SP6 phage RNA polymerase. In another embodiment, the molecule is synthesized by T3 phage RNA polymerase. In another embodiment, the molecule is synthesized by a polymerase selected from the above polymerases. In another embodiment, the RNA is synthesized chemically on a column similar to DNA. [0347] In another embodiment, the nucleoside that is modified in an RNA, oligoribonucleotide, or polyribonucleotide of the methods and compositions of the present disclosure is uridine (U). In another embodiment, the modified nucleoside is cytidine (C). In another embodiment, the modified nucleoside is adenine (A). In another embodiment the modified nucleoside is guanine (G). [0348] In another embodiment, the RNA of the methods and compositions of the present disclosure further comprises a poly-A tail. In another embodiment, the RNA of the methods and compositions of the present disclosure does not comprise a poly-A tail. Each possibility represents a separate embodiment of the present disclosure. [0349] In another embodiment, the RNA of the methods and compositions of the present disclosure comprises an m7GpppG cap. In another embodiment, the RNA of the methods and compositions of the present disclosure does not comprise an m7GpppG cap. In another embodiment, the RNA of the methods and compositions of the present disclosure comprises a 3′- O-methyl-m7GpppG. In another embodiment, the RNA of methods and composition of the present 270 11979815v1 P-627574-PC disclosure comprise a non-reversible cap analog, which, in some embodiments, is added during transcription of the RNA. In another embodiment, the RNA of methods and composition of the present disclosure comprise an anti-reverse cap analog. Each possibility represents a separate embodiment of the present disclosure. [0350] In another embodiment, the RNA of the methods and compositions of the present disclosure further comprises a cap-independent translational enhancer. In another embodiment, the RNA of the methods and compositions of the present disclosure does not comprise a cap- independent translational enhancer. In another embodiment, the cap-independent translational enhancer is a tobacco etch virus (TEV) cap-independent translational enhancer. In another embodiment, the cap-independent translational enhancer is any other cap-independent translational enhancer known in the art. Each possibility represents a separate embodiment of the present disclosure. [0351] In some embodiments, “pseudouridine” refers to m1acp3Ψ (1-methyl-3-(3-amino-5- carboxypropyl)pseudouridine. In another embodiment, the term refers to m1Ψ (1- methylpseudouridine). In another embodiment, the term refers to Ψm (2′-O-methylpseudouridine. In another embodiment, the term refers to m5D (5-methyldihydrouridine). In another embodiment, the term refers to m3Ψ (3-methylpseudouridine). In another embodiment, the modified nucleoside is 4' (pseudouridine). In another embodiment, the term refers to a pseudouridine moiety that is not further modified. In another embodiment, the term refers to a monophosphate, diphosphate, or triphosphate of any of the above pseudouridines. In another embodiment, the term refers to any other pseudouridine known in the art. Each possibility represents a separate embodiment of the present disclosure. [0352] In another embodiment, a modified RNA comprises a modified nucleoside, which in some embodiments, comprises m5C, m5U, m6A, s2U, ^ ^, 2'-O-methyl-U, 2’-O-methylpseudouridine, or a combination thereof. [0353] In another embodiment, the present disclosure provides a method for delivering a recombinant protein to a subject, the method comprising the step of contacting the subject with an RNA of the methods and compositions of the present disclosure, thereby delivering a recombinant protein to a subject. [0354] In another embodiment, a method of the present disclosure comprises increasing the number, percentage, or frequency of modified uridine nucleosides in the RNA molecule to decrease immunogenicity or increase efficiency of translation. In some embodiments, the number 271 11979815v1 P-627574-PC of modified uridine residues in an RNA, oligoribonucleotide, or polyribonucleotide molecule determines the magnitude of the effects observed in the present disclosure. [0355] In another embodiment, between 0.1% and 100% of the uridine residues in the modified RNAs of the methods and compositions of the present disclosure are modified (e.g. by the presence of pseudouridine). In another embodiment, 0.1% of the residues are modified. In another embodiment, 0.2%. In another embodiment, the fraction is 0.3%. In another embodiment, the fraction is 0.4%. In another embodiment, the fraction is 0.5%. In another embodiment, the fraction is 0.6%. In another embodiment, the fraction is 0.8%. In another embodiment, the fraction is 1%. In another embodiment, the fraction is 1.5%. In another embodiment, the fraction is 2%. In another embodiment, the fraction is 2.5%. In another embodiment, the fraction is 3%. In another embodiment, the fraction is 4%. In another embodiment, the fraction is 5%. In another embodiment, the fraction is 6%. In another embodiment, the fraction is 8%. In another embodiment, the fraction is 10%. In another embodiment, the fraction is 12%. In another embodiment, the fraction is 14%. In another embodiment, the fraction is 16%. In another embodiment, the fraction is 18%. In another embodiment, the fraction is 20%. In another embodiment, the fraction is 25%. In another embodiment, the fraction is 30%. In another embodiment, the fraction is 35%. In another embodiment, the fraction is 40%. In another embodiment, the fraction is 45%. In another embodiment, the fraction is 50%. In another embodiment, the fraction is 60%. In another embodiment, the fraction is 70%. In another embodiment, the fraction is 80%. In another embodiment, the fraction is 90%. In another embodiment, the fraction is 100%. [0356] In another embodiment, the fraction is less than 5%. In another embodiment, the fraction is less than 3%. In another embodiment, the fraction is less than 1%. In another embodiment, the fraction is less than 2%. In another embodiment, the fraction is less than 4%. In another embodiment, the fraction is less than 6%. In another embodiment, the fraction is less than 8%. In another embodiment, the fraction is less than 10%. In another embodiment, the fraction is less than 12%. In another embodiment, the fraction is less than 15%. In another embodiment, the fraction is less than 20%. In another embodiment, the fraction is less than 30%. In another embodiment, the fraction is less than 40%. In another embodiment, the fraction is less than 50%. In another embodiment, the fraction is less than 60%. In another embodiment, the fraction is less than 70% [0357] In another embodiment, 0.1% of the residues of a given uridine nucleotide are modified. In another embodiment, the fraction of the nucleotide is 0.2%. In another embodiment, the fraction 272 11979815v1 P-627574-PC is 0.3%. In another embodiment, the fraction is 0.4%. In another embodiment, the fraction is 0.5%. In another embodiment, the fraction is 0.6%. In another embodiment, the fraction is 0.8%. In another embodiment, the fraction is 1%. In another embodiment, the fraction is 1.5%. In another embodiment, the fraction is 2%. In another embodiment, the fraction is 2.5%. In another embodiment, the fraction is 3%. In another embodiment, the fraction is 4%. In another embodiment, the fraction is 5%. In another embodiment, the fraction is 6%. In another embodiment, the fraction is 8%. In another embodiment, the fraction is 10%. In another embodiment, the fraction is 12%. In another embodiment, the fraction is 14%. In another embodiment, the fraction is 16%. In another embodiment, the fraction is 18%. In another embodiment, the fraction is 20%. In another embodiment, the fraction is 25%. In another embodiment, the fraction is 30%. In another embodiment, the fraction is 35%. In another embodiment, the fraction is 40%. In another embodiment, the fraction is 45%. In another embodiment, the fraction is 50%. In another embodiment, the fraction is 60%. In another embodiment, the fraction is 70%. In another embodiment, the fraction is 80%. In another embodiment, the fraction is 90%. In another embodiment, the fraction is 100%. [0358] In another embodiment, the fraction of the given uridine nucleotide is less than 8%. In another embodiment, the fraction is less than 10%. In another embodiment, the fraction is less than 5%. In another embodiment, the fraction is less than 3%. In another embodiment, the fraction is less than 1%. In another embodiment, the fraction is less than 2%. In another embodiment, the fraction is less than 4%. In another embodiment, the fraction is less than 6%. In another embodiment, the fraction is less than 12%. In another embodiment, the fraction is less than 15%. In another embodiment, the fraction is less than 20%. In another embodiment, the fraction is less than 30%. In another embodiment, the fraction is less than 40%. In another embodiment, the fraction is less than 50%. In another embodiment, the fraction is less than 60%. In another embodiment, the fraction is less than 70%. [0359] In another embodiment, the terms “ribonucleotide,” “oligoribonucleotide,” and polyribonucleotide refers to, in some embodiments, compounds comprising nucleotides in which the sugar moiety is ribose. In another embodiment, the term includes both RNA and RNA derivates in which the backbone is modified. Numerous RNA backbone modifications are known in the art and contemplated in the present disclosure. In some embodiments, modified RNA is a PNA (peptide nucleic acid). PNA contain peptide backbones and nucleotide bases and are able to bind, in another embodiment, to both DNA and RNA molecules. In another embodiment, the nucleotide 273 11979815v1 P-627574-PC is modified by replacement of one or more phosphodiester bonds with a phosphorothioate bond. In another embodiment, the artificial nucleic acid contains any other variant of the phosphate backbone of native nucleic acids known in the art. Each nucleic acid derivative represents a separate embodiment of the present disclosure. [0360] Methods for production of nucleic acids having modified backbones are well known in the art, and are described, for example in U.S. Pat. Nos.5,723,335 and 5,663,153 issued to Hutcherson et al. and related PCT publication WO95/26204. Each method represents a separate embodiment of the present disclosure. [0361] The nucleic acid of interest can be purified by any method known in the art, or any method to be developed, so long as the method of purification removes contaminants from the nucleic acid preparation and thereby substantially reduces the immunogenicity potential of the nucleic acid preparation. In some embodiments, the nucleic acid of interest is purified using high-performance liquid chromatography (HPLC). In another embodiment, the nucleic acid of interest is purified by contacting the nucleic acid of interest with the bacterial enzyme RNase III. In other various embodiments, any method of nucleic acid purification that substantially reduces the immunogenicity of the nucleic acid preparation can be used. Non-limiting examples of purification methods that can be used with the compositions and methods of the disclosure include liquid chromatography separation and enzyme digestion, each used alone or in any combination, simultaneously or in any order. Non-limiting examples of liquid chromatography separation include HPLC and fast protein liquid chromatography (FPLC). Materials useful in the HPLC and FPLC methods of the disclosure include, but are not limited to, cross-linked polystyrene/divinylbenzene (PS/DVB), PS/DVB-C18, PS/DVB-alkylated, Helix DNA columns (Varian), Eclipse dsDNA Analysis Columns (Agilent Technologies), Reverse-phase 5 (RPC-5) exchange material, DNAPac, ProSwift, and bio-inert UltiMate.RTM. 3000 Titanium columns (Dionex). Enzymes useful in the enzyme digestion methods of the disclosure include any enzyme able to digest any contaminant in a nucleic acid preparation of the disclosure, such as, for example a dsRNA contaminant, and include but are not limited to, RNase III, RNase V1, Dicer, and Chipper (see Fruscoloni et al., 2002, PNAS 100:1639) Non-limiting examples of assays for assessing the purity of the nucleic acid of interest include a dot-blot assay, a Northern blot assay, and a dendritic cell activation assay, as described elsewhere herein. [0362] In another embodiment, the modified RNA of the methods and compositions of the present disclosure is significantly less immunogenic than an unmodified in vitro-synthesized RNA 274 11979815v1 P-627574-PC molecule with the same sequence. In another embodiment, the modified RNA molecule is 2-fold less immunogenic than its unmodified counterpart. In another embodiment, immunogenicity is reduced by a 3-fold factor. In another embodiment, immunogenicity is reduced by a 5-fold factor. In another embodiment, immunogenicity is reduced by a 7-fold factor. In another embodiment, immunogenicity is reduced by a 10-fold factor. In another embodiment, immunogenicity is reduced by a 15-fold factor. In another embodiment, immunogenicity is reduced by a fold factor. In another embodiment, immunogenicity is reduced by a 50-fold factor. In another embodiment, immunogenicity is reduced by a 100-fold factor. In another embodiment, immunogenicity is reduced by a 200-fold factor. In another embodiment, immunogenicity is reduced by a 500-fold factor. In another embodiment, immunogenicity is reduced by a 1000-fold factor. In another embodiment, immunogenicity is reduced by a 2000-fold factor. In another embodiment, immunogenicity is reduced by another fold difference. [0363] In another embodiment, “significantly less immunogenic” refers to a detectable decrease in immunogenicity. In another embodiment, the term refers to a fold decrease in immunogenicity (e.g. 1 of the fold decreases enumerated above). In another embodiment, the term refers to a decrease such that an effective amount of the modified RNA can be administered without triggering a detectable immune response. In another embodiment, the term refers to a decrease such that the modified RNA can be repeatedly administered without eliciting an immune response sufficient to detectably reduce expression of the recombinant protein. In another embodiment, the decrease is such that the modified RNA can be repeatedly administered without eliciting an immune response sufficient to eliminate detectable expression of the recombinant protein. [0364] Methods of determining immunogenicity are well known in the art, and described in detail in U. S. Patent 8,278,036 which is hereby incorporated by reference herein. [0365] In another embodiment, the modified RNA of the methods and compositions of the present disclosure is translated in the cell more efficiently than an unmodified RNA molecule with the same sequence. In another embodiment, the modified RNA exhibits enhanced ability to be translated by a target cell. In another embodiment, translation is enhanced by a factor of 2-fold relative to its unmodified counterpart. In another embodiment, translation is enhanced by a 3-fold factor. In another embodiment, translation is enhanced by a 5-fold factor. In another embodiment, translation is enhanced by a 7-fold factor. In another embodiment, translation is enhanced by a 10- fold factor. In another embodiment, translation is enhanced by a 15-fold factor. In another embodiment, translation is enhanced by a 20-fold factor. In another embodiment, translation is 275 11979815v1 P-627574-PC enhanced by a 50-fold factor. In another embodiment, translation is enhanced by a 100-fold factor. In another embodiment, translation is enhanced by a 200-fold factor. In another embodiment, translation is enhanced by a 500-fold factor. In another embodiment, translation is enhanced by a 1000-fold factor. In another embodiment, translation is enhanced by a 2000-fold factor. In another embodiment, the factor is 10-1000-fold. In another embodiment, the factor is 10-100-fold. In another embodiment, the factor is 10-200-fold. In another embodiment, the factor is 10-300-fold. In another embodiment, the factor is 10-500-fold. In another embodiment, the factor is 20-1000- fold. In another embodiment, the factor is 30-1000-fold. In another embodiment, the factor is 50- 1000-fold. In another embodiment, the factor is 100-1000-fold. In another embodiment, the factor is 200-1000-fold. In another embodiment, translation is enhanced by any other significant amount or range of amounts. Each possibility represents a separate embodiment of the present disclosure. [0366] Methods of determining translation efficiency are well known in the art, and include, e.g. measuring the activity of an encoded reporter protein (e.g., luciferase or renilla or green fluorescent protein [Wall A A, Phillips A M et al, Effective translation of the second cistron in two Drosophila dicistronic transcripts is determined by the absence of in-frame AUG codons in the first cistron. J Biol Chem 2005; 280(30): 27670-8]), or measuring radioactive label incorporated into the translated protein (Ngosuwan J, Wang N M et al., Roles of cytosolic Hsp70 and Hsp40 molecular chaperones in post-translational translocation of pre-secretory proteins into the endoplasmic reticulum. J Biol Chem 2003; 278(9): 7034-42). Each method represents a separate embodiment of the present disclosure. [0367] In another embodiment, the target cell of the method of the present disclosure is a dendritic cell. In another embodiment, the target cell of the method of the present disclosure is a macrophage. In another embodiment, the target cell of the method of the present disclosure is a B cell. In another embodiment, the target cell of the method of the present disclosure is another antigen presenting cell. In another embodiment, the target cell of methods of the present disclosure is a mucosal cell. In another embodiment, the target cell of methods of the present disclosure is an epithelial cell. In another embodiment, the cell is a skin cell. In another embodiment, the cell is an epidermal cell. In another embodiment, the cell is a keratinocyte. In another embodiment, the cell is a Merkel cell, melanocyte or Langerhans cell. Each possibility represents a separate embodiment of the present disclosure. C. Codon Optimization and GC Enrichment [0368] The present disclosure also provides codon optimized nucleotide sequences. 276 11979815v1 P-627574-PC [0369] As used herein, the term “codon-optimized” refers to alteration of codons in a coding region of a nucleic acid molecule (e.g., a nucleotide sequence) to reflect the typical codon usage of a host organism (e.g., a subject receiving a nucleic acid molecule (e.g., a nucleotide sequence)) preferably without altering the amino acid sequence encoded by the nucleic acid molecule. Within the context of the present disclosure, in some embodiments, coding regions are codon-optimized for optimal expression in a subject to be treated using the RNA molecules described herein. In some embodiments, codon-optimization may be performed such that codons for which frequently occurring tRNAs are available are inserted in place of “rare codons.” In some embodiments, codon-optimization may include increasing guanosine/cytosine (G/C) content of a coding region of RNA described herein as compared to the G/C content of the corresponding coding sequence of a wild type RNA, wherein the amino acid sequence encoded by the RNA is preferably not modified compared to the amino acid sequence. [0370] In some embodiments, a coding sequence (also referred to as a “coding region”) is codon optimized for expression in the subject to whom a composition (e.g., a pharmaceutical composition) is to be administered (e.g., a human). Thus, in some embodiments, sequences in such a polynucleotide (e.g., a nucleotide sequence) may differ from wild type sequences encoding the relevant antigen, fragment or epitope thereof, even when the amino acid sequence of the antigen, fragment or epitope thereof is wild type. [0371] In some embodiments, strategies for codon optimization for expression in a relevant subject (e.g., a human), and even, in some cases, for expression in a particular cell or tissue. [0372] Various species exhibit particular bias for certain codons of a particular amino acid. Without wishing to be bound by any one theory, codon bias (differences in codon usage between organisms) often correlates with the efficiency of translation of messenger RNA (mRNA), which is in turn believed to be dependent on, among other things, the properties of the codons being translated and the availability of particular transfer RNA (tRNA) molecules. The predominance of selected tRNAs in a cell may generally be a reflection of the codons used most frequently in peptide synthesis. Accordingly, genes may be tailored for optimal gene expression in a given organism based on codon optimization. Codon usage tables are available, for example, at the "Codon Usage Database" available at www.kazusa.orjp/codon/ and these tables may be adapted in a number of ways. Computer algorithms for codon optimizing a particular sequence for expression in a particular subject or its cells are also available, such as Gene Forge (Aptagen; Jacobus, PA), are also available. 277 11979815v1 P-627574-PC [0373] In some embodiments, a polynucleotide (e.g., a polyribonucleotide or a nucleotide sequence) of the present disclosure is codon optimized, wherein the codons in the polynucleotide (e.g., the polyribonucleotide) are adapted to human codon usage (herein referred to as “human codon optimized polynucleotide”). In some embodiments, a portion of a nucleotide sequence is codon optimized (e.g., a portion of or the portion encoding a glycoprotein or a portion of or the portion encoding a signal sequence). In some embodiments, the entire nucleotide sequence is codon optimized. Codons encoding the same amino acid occur at different frequencies in a subject, e.g., a human. Accordingly, in some embodiments, the coding sequence of a polynucleotide of the present disclosure is modified such that the frequency of the codons encoding the same amino acid corresponds to the naturally occurring frequency of that codon according to the human codon usage, e.g., as shown in Table 7. For example, in the case of the amino acid Ala, the wild type coding sequence is preferably adapted in a way that the codon “GCC” is used with a frequency of 0.40, the codon “GCT” is used with a frequency of 0.28, the codon “GCA” is used with a frequency of 0.22 and the codon “GCG” is used with 30 a frequency of 0.10 etc. (see Table 7). Accordingly, in some embodiments, such a procedure (as exemplified for Ala) is applied for each amino acid encoded by the coding sequence of a polynucleotide to obtain sequences adapted to human codon usage. Table 7: Human codon usage with frequencies indicated for each amino acid. Amino Codon Frequency Amino Codon Frequency
Figure imgf000280_0001
11979815v1 P-627574-PC Amino Codon Frequency Amino Codon Frequency Acid Acid
Figure imgf000281_0001
[0374] Certain strategies for codon optimization and/or G/C enrichment for human expression are described in WO2002/098443, which is incorporated by reference herein in its entirety. In some embodiments, a coding sequence may be optimized using a multiparametric optimization strategy. In some embodiments, optimization parameters may include parameters that influence protein expression, which can be, for example, impacted on a transcription level, an RNA level, and/or a translational level. In some embodiments, exemplary optimization parameters include, but are not limited to transcription-level parameters (including, e.g., GC content, consensus splice sites, cryptic splice sites, SD sequences, TATA boxes, termination signals, artificial recombination sites, and combinations thereof); RNA-level parameters (including, e.g., RNA instability motifs, 279 11979815v1 P-627574-PC ribosomal entry sites, repetitive sequences, and combinations thereof); translation-level parameters (including, e.g., codon usage, premature poly(A) sites, ribosomal entry sites, secondary structures, and combinations thereof); or combinations thereof. In some embodiments, a coding sequence may be optimized by a GeneOptimizer algorithm as described in Fath et al. “Multiparameter RNA and Codon Optimization: A Standardized Tool to Assess and Enhance Autologous Mammalian Gene Expression” PLoS ONE 6(3): e17596; Rabb et al., which is incorporated herein by reference in its entirety, “The GeneOptimizer Algorithm: using a sliding window approach to cope with the vast sequence space in multiparameter DNA sequence optimization” Systems and Synthetic Biology (2010) 4:215-225; and Graft et al. “Codon- optimized genes that enable increased heterologous expression in mammalian cells and elicit efficient immune responses in mice after vaccination of naked DNA” Methods Mol Med (2004) 94:197-210, the entire content of each of which is incorporated herein for the purposes described herein. In some embodiments, a coding sequence may be optimized by Eurofins’ adaption and optimization algorithm “GENEius” as described in Eurofins’ Application Notes: Eurofins’ adaption and optimization software “GENEius” in comparison to other optimization algorithms, the entire content of which is incorporated by reference for the purposes described herein. [0375] In some embodiments, a coding sequence utilized in accordance with the present disclosure has G/C content that is increased compared to a coding sequence for an HSV gC, gD, and/or gE (or immunogenic fragment thereof) construct described herein. In some embodiments, guanosine/cytidine (G/C) content of a coding region is modified relative to a comparable coding sequence for an HSV gC, gD, and/or gE (or immunogenic fragment thereof) construct described herein, but the amino acid sequence encoded by the nucleotide sequence is not modified. [0376] Without wishing to be bound by any particular theory, it is proposed that GC enrichment may improve translation of a payload sequence. Typically, sequences having an increased G (guanosine)/C (cytidine) content are more stable than sequences having an increased A (adenosine)/U (uridine) content. In respect to the fact that several codons code for one and the same amino acid (so-called degeneration of the genetic code), the most favorable codons for the stability can be determined (so-called alternative codon usage). Depending on the amino acid to be encoded by a nucleotide sequence, there are various possibilities for modification of the ribonucleic acid sequence, compared to its wild type sequence. In particular, codons which contain A and/or U nucleosides can be modified by substituting these codons by other codons, which code 280 11979815v1 P-627574-PC for the same amino acids but contain no A and/or U or contain a lower content of A and/or U nucleosides. [0377] In some embodiments, G/C content of a coding region of a nucleotide sequence described herein is increased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G/C content of the coding region prior to codon optimization, e.g., of the wild type RNA. In some embodiments, G/C content of a coding region of a nucleotide sequence described herein is decreased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G/C content of the coding region prior to codon optimization, e.g., of the wild type RNA. [0378] In some embodiments, stability and translation efficiency of a nucleotide sequence may incorporate one or more elements established to contribute to stability and/or translation efficiency of the nucleotide sequence; exemplary such elements are described, for example, in PCT/EP2006/009448 incorporated herein by reference. In some embodiments, to increase expression of a nucleotide sequence used according to the present disclosure, a nucleotide sequence may be modified within the coding region, i.e., the sequence encoding the expressed peptide or protein, without altering the sequence of the expressed peptide or protein, for example so as to increase the GC-content to increase RNA stability and/or to perform a codon optimization and, thus, enhance translation in cells. Methods of Treatment and Uses of the Compositions [0379] In some embodiments, the present disclosure provides methods of inhibiting an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of suppressing an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of preventing an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating an HSV oral mucosal infection in a subject comprising administering a composition of the present disclosure. [0380] In some embodiments, an HSV oral mucosal infection is an HSV-1 infection. In another embodiment, an oral mucosal infection is an HSV-2 infection. In other embodiments, the present 281 11979815v1 P-627574-PC disclosure provides methods of treating an oral mucosal infection in a subject comprising administering a composition of the present disclosure. [0381] In some embodiments, the present disclosure provides methods of inhibiting an HSV-1 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of suppressing an HSV- 1 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of an HSV-1 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of preventing an HSV-1 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating an HSV-1 oral mucosal infection in a subject comprising administering a composition of the present disclosure. [0382] In some embodiments, the present disclosure provides methods of inhibiting an HSV-2 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of suppressing an HSV- 2 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of an HSV-2 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of preventing an HSV-2 oral mucosal infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating an HSV-2 oral mucosal infection in a subject comprising administering a composition of the present disclosure. [0383] In some embodiments, the present disclosure provides methods of inhibiting HSV encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of suppressing HSV encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of HSV encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of preventing HSV encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present 282 11979815v1 P-627574-PC disclosure provides methods of treating HSV encephalitis in a subject comprising administering a composition of the present disclosure. [0384] In some embodiments, HSV encephalitis is an HSV-1 infection. In another embodiment, an oral mucosal infection is an HSV-2 infection. In other embodiments, the present disclosure provides methods of treating an oral mucosal infection in a subject comprising administering a composition of the present disclosure. [0385] In some embodiments, the present disclosure provides methods of inhibiting HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of suppressing HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of preventing HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating HSV-1 encephalitis in a subject comprising administering a composition of the present disclosure. [0386] In some embodiments, the present disclosure provides methods of inhibiting HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of suppressing HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of reducing the incidence of HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of preventing HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating HSV-2 encephalitis in a subject comprising administering a composition of the present disclosure. [0387] In other embodiments, the present disclosure provides methods of preventing establishment of a latent HSV infection in a subject comprising administering a composition of the present disclosure. In other embodiments, the present disclosure provides methods of treating a latent HSV infection in a subject comprising administering a composition of the present disclosure. In some embodiments, the latent HSV infection is an HSV-1 infection. In other embodiments, the latent HSV infection is an HSV-2 infection. 283 11979815v1 P-627574-PC [0388] In some embodiments, the latent HSV infection is in the dorsal root ganglia. In other embodiments, the latent HSV infection is in the trigeminal ganglia. In other embodiments, the latent HSV infection is in the brain. In other embodiments, the latent HSV infection is in the olfactory bulb. In other embodiments, the latent HSV infection is in any combination of dorsal root ganglia, trigeminal ganglia, brain, or olfactory bulb. [0389] In some embodiments, the latent HSV infection is detected in the dorsal root ganglia. In other embodiments, the latent HSV infection is detected in the trigeminal ganglia. In other embodiments, the latent HSV infection is detected in the brain. In other embodiments, the latent HSV infection is detected in the olfactory bulb. In other embodiments, the latent HSV infection is detected in any combination of dorsal root ganglia, trigeminal ganglia, brain, or olfactory bulb. In some embodiments, the latent HSV infection is an HSV-1 infection. In other embodiments, the latent HSV infection is an HSV-2 infection. [0390] In other embodiments, the present disclosure provides methods of suppressing or inhibiting an HSV infection of the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or a combination thereof in a subject comprising administering to said subject a composition of the present disclosure. [0391] In other embodiments, the present disclosure provides methods of treating or inhibiting an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof. In some embodiments, the HSV-2 protein comprises an HSV-2 glycoprotein, or an immunogenic fragment thereof. In some embodiments, the HSV-2 glycoprotein comprises an HSV-2 glycoprotein D (gD) or an immunogenic fragment thereof, an HSV glycoprotein C (gC) or an immunogenic fragment thereof, and an HSV glycoprotein E (gE) or an immunogenic fragment thereof. For example, said RNAs can be one or more RNAs comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6. [0392] In other embodiments, the present disclosure provides methods of inhibiting an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof. In other embodiments, the present disclosure provides methods of treating an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding 284 11979815v1 P-627574-PC Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof. In other embodiments, the present disclosure provides methods of suppressing an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof. In other embodiments, the present disclosure provides methods of reducing the incidence of an HSV-1 infection in a subject comprising the step of administering a composition comprising one or more RNA encoding Herpes Simplex Virus-2 (HSV-2) proteins, or immunogenic fragments thereof. For example, said RNAs can be one or more RNAs comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6. [0393] In some embodiments, the present disclosure provides methods of vaccinating a subject against HSV and treating, impeding, inhibiting, reducing the incidence of, or suppressing an HSV infection or a symptom or manifestation thereof, comprising administration of a composition of the present disclosure. [0394] In some embodiments, the present disclosure provides a method for treating an HSV infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof. [0395] In another embodiment, the present disclosure provides a method for suppressing an HSV infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof. [0396] In another embodiment, the present disclosure provides a method for inhibiting an HSV infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof. [0397] In another embodiment, the present disclosure provides a method for reducing the incidence of HSV infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV glycoprotein or immunogenic fragment thereof. 285 11979815v1 P-627574-PC [0398] In some embodiments, the HSV infection is an HSV-1 infection. In another embodiment, the HSV infection is an HSV-2 infection. [0399] In some embodiments, the subject is administered HSV-1 glycoproteins, or immunogenic fragments thereof, for methods of treating, inhibiting, suppressing, etc. an HSV-1 infection. In another embodiment, the subject is administered HSV-2 glycoproteins, or immunogenic fragments thereof, for methods of treating, inhibiting, suppressing, etc. an HSV-2 infection. In another embodiment, the subject is administered HSV-1 glycoproteins, or immunogenic fragments thereof, for methods of treating, inhibiting, suppressing, etc. an HSV-1 infection, HSV-2 infection, or a combination thereof. In another embodiment, the subject is administered HSV-2 glycoproteins, or immunogenic fragments thereof, for methods of treating, inhibiting, suppressing, etc. an HSV-1 infection, HSV-2 infection, or a combination thereof. In some embodiments, administration of HSV-1 glycoproteins (e.g., gC1, gD1, gE1, immunogenic fragments thereof or a combination thereof) treats or prevents HSV-1 and HSV-2 infection. In another embodiment, administration of HSV-2 glycoproteins (e.g., gC2, gD2, gE2, immunogenic fragments thereof, or a combination thereof) treats or prevents HSV-1 and HSV-2 infection. [0400] According to this aspect and in some embodiments, the present disclosure provides a method for treating, suppressing, inhibiting, or reducing the incidence of Herpes Simplex Virus 1 (HSV-1) infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV-1 glycoprotein or immunogenic fragment thereof. [0401] In some embodiments, the present disclosure provides a method for treating, suppressing, inhibiting, or reducing the incidence of Herpes Simplex Virus 2 (HSV-2) infection in a subject, comprising contacting said subject with a composition comprising one or more RNAs, wherein each of said RNAs encodes an HSV-2 glycoprotein or immunogenic fragment thereof. [0402] In some embodiments, said contacting is via administration to said subject. [0403] In another embodiment, the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV gD or immunogenic fragment thereof; (b) an HSV gC or immunogenic fragment thereof as described herein; (c) an HSV gE or immunogenic fragment thereof as described herein, or a combination thereof. 286 11979815v1 P-627574-PC [0404] In another embodiment, the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV-2 gD or immunogenic fragment thereof; (b) an HSV-2 gC or immunogenic fragment thereof as described herein; and (c) an HSV-2 gE or immunogenic fragment thereof as described herein, or a combination thereof. [0405] In another embodiment, the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV-1 gD or immunogenic fragment thereof; (b) an HSV-1 gC or immunogenic fragment thereof as described herein; and (c) an HSV-1 gE or immunogenic fragment thereof as described herein, or a combination thereof. [0406] In another embodiment, the present disclosure provides a method of inducing an anti-HSV immune response in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV gD or immunogenic fragment thereof; (b) an HSV gC or immunogenic fragment thereof as described herein; (c) an HSV gE or immunogenic fragment thereof as described herein, or a combination thereof. [0407] In another embodiment, the present disclosure provides a method of inducing an anti-HSV immune response in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV-2 gD or immunogenic fragment thereof; (b) an HSV-2 gC or immunogenic fragment thereof as described herein; and (c) an HSV-2 gE or immunogenic fragment thereof as described herein, or a combination thereof. [0408] In another embodiment, the present disclosure provides a method of inducing an anti-HSV immune response in a subject, the method comprising the step of administering to said subject an immunogenic composition comprising RNAs encoding: (a) an HSV-1 gD or immunogenic fragment thereof; (b) an HSV-1 gC or immunogenic fragment thereof as described herein; and (c) an HSV-1 gE or immunogenic fragment thereof as described herein, or a combination thereof. [0409] In another embodiment, the present disclosure provides a method of inhibiting a primary HSV infection in a subject, the method comprising the step of administering to the subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of treating an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present 287 11979815v1 P-627574-PC disclosure provides a method of reducing the incidence of an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of inhibiting a flare following a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0410] In some embodiments, the present disclosure provides methods of treating and/or suppressing a primary HSV infection and/or a secondary HSV infection. In some embodiments, a “primary” infection refers to a first-time infection. In some embodiments, a “secondary” infection refers to a recurrence of an HSV infection. [0411] In some embodiments, a “flare” or “recurrence” refers to reinfection of skin tissue following latent neuronal HSV infection. In another embodiment, the terms refer to reactivation of HSV after a latency period. In another embodiment, the terms refer to symptomatic HSV lesions following a non-symptomatic latency period. [0412] In another embodiment, the present disclosure provides a method of inhibiting spread of HSV. In some embodiments, the spread from DRG to skin is inhibited. In some embodiments, cell-to-cell spread of HSV is inhibited. In some embodiments, anterograde spread is inhibited. In some embodiments, retrograde spread is inhibited. “DRG” refers, in some embodiments, to a neuronal cell body and in another embodiment, contain the neuron cell bodies of nerve fibers. In another embodiment, the term refers to any other definition of “DRG” used in the art. In another embodiment, spread of HSV to neural tissue is inhibited. [0413] In another embodiment, the present disclosure provides a method of inhibiting a recurrence following a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of preventing a recurrence following a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0414] In another embodiment, the present disclosure provides a method of inhibiting an HSV labialis following a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0415] In another embodiment, the present disclosure provides a method of preventing a recurrence of an HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a 288 11979815v1 P-627574-PC method of diminishing the severity of a recurrence of an HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of reducing the frequency of a recurrence of an HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure. In some embodiments, the present disclosure provides any of the described methods in an HIV-infected subject. [0416] In another embodiment, the present disclosure provides a method of treating HSV encephalitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of reducing the incidence of HSV encephalitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. “HSV encephalitis” refers, In some embodiments, to encephalitis caused by a Herpes Simplex Virus-1 (HSV). In another embodiment, the term refers to encephalitis associated with HSV. In another embodiment, the term refers to any other type of HSV-mediated encephalitis known in the art. [0417] In another embodiment, the present disclosure provides a method of treating or reducing an HSV neonatal infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0418] In another embodiment, the present disclosure provides a method for introducing an HSV glycoprotein to a cell of a subject, comprising contacting said cell with an in vitro-transcribed RNA molecule encoding the recombinant protein, wherein said in vitro-transcribed RNA molecule further comprises a modified nucleoside, thereby introducing said HSV glycoprotein, or immunogenic fragment thereof, into said cell of said subject. [0419] In another embodiment, the present disclosure provides a method for inducing a mammalian cell to produce an HSV glycoprotein, or immunogenic fragment thereof, comprising contacting said mammalian cell with an in vitro-synthesized RNA molecule encoding the HSV glycoprotein, or immunogenic fragment thereof, the in vitro-synthesized RNA molecule comprising a pseudouridine, thereby inducing said mammalian cell to produce said HSV glycoprotein, or immunogenic fragment thereof. [0420] It is to be understood that reference to HSV herein refers in some embodiments, to HSV- 1, while in another embodiment, to HSV-2, while in another embodiment, to HSV-1 and HSV-2. [0421] In some embodiments, “HSV-1” refers to a Herpes Simplex Virus-1. In some embodiments, “HSV-1” refers to a HSV-1 strain. In another embodiment, the term refers to a KOS 289 11979815v1 P-627574-PC strain. In another embodiment, the term refers to an F strain. In another embodiment, the term refers to an NS strain. In another embodiment, the term refers to a CL101 strain. In another embodiment, the term refers to a “17” strain. In another embodiment, the term refers to a “17+syn” strain. In another embodiment, the term refers to a MacIntyre strain. In another embodiment, the term refers to an MP strain. In another embodiment, the term refers to an HF strain. In another embodiment, the term refers to any other HSV-1 strain known in the art. [0422] In some embodiments,“HSV-2” refers to a Herpes Simplex Virus-2. In some embodiments,“HSV-2” refers to a HSV-2 strain. In another embodiment, the term refers to an HSV-2333 strain. In another embodiment, the term refers to a 2.12 strain. In another embodiment, the term refers to an HG52 strain. In another embodiment, the term refers to an MS strain. In another embodiment, the term refers to a G strain. In another embodiment, the term refers to a 186 strain. In another embodiment, the term refers to any other HSV-2 strain known in the art. [0423] In another embodiment, the present disclosure provides a method of vaccinating a subject against an HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of suppressing an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of impeding an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of impeding a primary HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of impeding neuronal HSV spread in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0424] The terms “impeding an HSV infection” and “impeding a primary HSV infection” refer, in another embodiment, to decreasing the titer of infectious virus. In another embodiment, the terms refer to decreasing the extent of viral replication. [0425] In another embodiment, the present disclosure provides a method of treating an HSV- mediated herpetic ocular disease in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of reducing the incidence of an HSV-mediated herpetic ocular disease in a subject, the method comprising the step of administering to said subject a composition of the 290 11979815v1 P-627574-PC present disclosure. In some embodiments, the herpetic ocular disease comprises a corneal infection. In another embodiment, the present disclosure provides a method of treating an HSV-1 corneal infection or herpes keratitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of reducing the incidence of an HSV-1 corneal infection or herpes keratitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In some embodiments, the HSV-1 corneal infection or herpes keratitis is an HSV-1 corneal infection or herpes keratitis. [0426] In another embodiment, the present disclosure provides a method of treating herpetic stomatitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of reducing the incidence of herpetic stomatitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In some embodiments, the stomatitis is an HSV-1 stomatitis. [0427] In another embodiment, the present disclosure provides a method of treating, suppressing or inhibiting an HSV genital infection, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of treating, suppressing or inhibiting any manifestation of recurrent HSV infection, the method comprising the step of administering to said subject a composition of the present disclosure. [0428] In another embodiment, the present disclosure provides a method of reducing the incidence of an HSV-mediated genital ulcer disease in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0429] In another embodiment, the present disclosure provides a method of impeding establishment of a latent HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of preventing establishment of a latent HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of suppressing the establishment of a latent HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. In some embodiments, establishment of a latent HSV infection in a subject is through HSV infection of the 291 11979815v1 P-627574-PC DRG. In another embodiment, establishment of a latent HSV infection in a subject is through HSV infection of the trigeminal nerve or trigeminal ganglia. [0430] In some embodiments, the present disclosure provides a method of treating, suppressing or inhibiting a genital herpes infection in a subject, comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of treating, suppressing or inhibiting an oral herpes infection in a subject, comprising the step of administering to said subject a composition of the present disclosure. [0431] In another embodiment, the present disclosure provides a method of treating or inhibiting an orolabial herpes infection in a subject, comprising the step of administering to said subject a composition of the present disclosure. In some embodiment, the HSV infection is on a lip of the subject. [0432] In another embodiment, the present disclosure provides a method of reducing the incidence of an HSV-mediated encephalitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0433] In another embodiment, the herpes-mediated encephalitis treated or prevented by a method of the present disclosure is a focal herpes encephalitis. In another embodiment, the herpes- mediated encephalitis is a neonatal herpes encephalitis. In another embodiment, the herpes- mediated encephalitis is any other type of herpes-mediated encephalitis known in the art. [0434] In another embodiment, the present disclosure provides a method of treating or reducing the incidence of a disease, disorder, or symptom associated with or secondary to an HSV-mediated encephalitis in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0435] In another embodiment, the present disclosure provides a method of treating, reducing the pathogenesis of, ameliorating the symptoms of, ameliorating the secondary symptoms of, reducing the incidence of, prolonging the latency to a relapse of an HSV infection in a subject, comprising the step of administering to the subject a composition of the present disclosure. [0436] In another embodiment, the present disclosure provides a method of protecting a subject against formation of a zosteriform lesion or an analogous outbreak in a human subject. In another embodiment, the present disclosure provides a method of inhibiting the formation of an HSV zosteriform lesion or an analogous outbreak in a human subject. [0437] “Zosteriform” refers, in some embodiments, to skin lesions characteristic of an HSV infection, particularly during reactivation infection, which, in some embodiments, begin as a rash 292 11979815v1 P-627574-PC and follow a distribution near dermatomes, commonly occurring in a strip or belt-like pattern. In some embodiments, the rash evolves into vesicles or small blisters filled with serous fluid. In some embodiments, zosteriform lesions form in mice as a result of contact with HSV. In another embodiment, zosteriform lesions form in humans as a result of contact with HSV. “Zosteriform spread” refers, in some embodiments, to an HSV infection that spreads from the ganglia to secondary skin sites within the dermatome. In another embodiment, the term refers to spread within the same dermatome as the initial site of infection. In another embodiment, the term refers to any other definition of “zosteriform spread” known in the art. “Outbreak”, in another embodiment, refers to a sudden increase in symptoms of a disease or in the spread or prevalence of a disease, and in some embodiments, refers to a sudden increase in zosteriform lesions, while in another embodiment, “outbreak” refers to a sudden eruption of zosteriform lesions. [0438] In some embodiments, the present disclosure provides a method of impeding the formation of a dermatome lesion or an analogous condition in a subject. In some embodiments, dermatome lesions form as a result of contact with HSV. In another embodiment, dermatome lesions most often develop when the virus reactivates from latency in the ganglia and in some embodiments, spreads down nerves, in some embodiments, causing a recurrent infection. [0439] It is to be understood that the methods of the present disclosure may be used to treat, inhibit, suppress, etc. an HSV infection or primary or secondary symptoms related to such an infection following exposure of the subject to HSV. In another embodiment, the subject has been infected with HSV before vaccination. In another embodiment, the subject is at risk for HSV infection. In another embodiment, whether or not the subject has been infected with HSV at the time of vaccination, vaccination by a method of the present disclosure is efficacious in treating, inhibiting, suppressing, etc. an HSV infection or primary or secondary symptoms related to such an infection. [0440] In some embodiments, “treating” refers to either therapeutic treatment or prophylactic or preventative measures, wherein the object is to prevent or lessen the targeted pathologic condition or disorder as described hereinabove. Thus, in some embodiments, treating may include directly affecting or curing, suppressing, inhibiting, preventing, reducing the severity of, delaying the onset of, reducing symptoms associated with the disease, disorder or condition, or a combination thereof. Thus, in some embodiments, “treating” refers inter alia to delaying progression, expediting remission, inducing remission, augmenting remission, speeding recovery, increasing efficacy of or decreasing resistance to alternative therapeutics, or a combination thereof. In some embodiments, “preventing” refers, inter alia, to delaying the onset of symptoms, preventing 293 11979815v1 P-627574-PC relapse to a disease, decreasing the number or frequency of relapse episodes, increasing latency between symptomatic episodes, or a combination thereof. In some embodiments, “suppressing” or “inhibiting”, refers inter alia to reducing the severity of symptoms, reducing the severity of an acute episode, reducing the number of symptoms, reducing the incidence of disease-related symptoms, reducing the latency of symptoms, ameliorating symptoms, reducing secondary symptoms, reducing secondary infections, prolonging patient survival, or a combination thereof. [0441] In some embodiments, the compositions and methods of the present disclosure are effective in lowering HSV acquisition rates, duration of HSV infection, frequency of HSV reactivation, or a combination thereof. In another embodiment, the compositions and methods of the present disclosure are effective in treating or inhibiting genital ulcer disease, which in some embodiments, entails decreasing the severity or frequency of HSV genital ulcer disease. In some embodiments, the compositions and methods of the present disclosure block immune evasion from complement. In some embodiments, vaccination with RNA-encoded HSV subunits, or immunogenic fragments thereof, may produce high titers of neutralizing antibodies or potent T-cell responses; however, upon subsequent infection, HSV immune evasion molecules may block the activities of antibodies or T cells, thereby reducing composition efficacy. In some embodiments, the compositions and methods of the present disclosure incorporate strategies to block virus mediated immune evasion by, in some embodiments, enhancing the effectiveness of e.g. a gD-1 subunit, or immunogenic fragment thereof, composition using gC-1, or immunogenic fragments thereof, to prevent immune evasion from complement. [0442] In some embodiments, studies in guinea pigs and mice suggest that viral load in ganglia correlates with the frequency of recurrent HSV infections. Thus, in some embodiments, the compositions and methods of the present disclosure are useful for preventing or inhibiting recurrent HSV infections. In some embodiments, antibodies to e.g. gC-1 block domains involved in immune evasion, which enhances complement activity, improves neutralizing activity of anti- gD-1 IgG, increases antibody- and complement-dependent cellular cytotoxicity, and augments complement-mediated neutralization and lysis of infected cells. [0443] In some embodiments, symptoms are primary, while in another embodiment, symptoms are secondary. In some embodiments, “primary” refers to a symptom that is a direct result of the subject viral infection, while in some embodiments, “secondary” refers to a symptom that is derived from or consequent to a primary cause. In some embodiments, the compositions and strains 294 11979815v1 P-627574-PC for use in the present disclosure treat primary or secondary symptoms or secondary complications related to HSV infection. [0444] In another embodiment, “symptoms” may be any manifestation of an HSV infection, comprising blisters, ulcerations, or lesions on the urethra, cervix, upper thigh, and/or anus in women and on the penis, urethra, scrotum, upper thigh, and anus in men, inflammation, swelling, fever, flu-like symptoms, sore mouth, sore throat, pharyngitis, pain, blisters on tongue, mouth or lips, ulcers, cold sores, neck pain, enlarged lymph nodes, reddening, bleeding, itching, dysuria, headache, muscle pain, etc., or a combination thereof. [0445] In another embodiment, the disease, disorder, or symptom is fever. In another embodiment, the disease, disorder, or symptom is headache. In another embodiment, the disease, disorder, or symptom is stiff neck. In another embodiment, the disease, disorder, or symptom is seizures. In another embodiment, the disease, disorder, or symptom is partial paralysis. In another embodiment, the disease, disorder, or symptom is stupor. In another embodiment, the disease, disorder, or symptom is coma. In another embodiment, the disease, disorder, or symptom is any other disease, disorder, or symptom known in the art that is associated with or secondary to a herpes-mediated encephalitis. [0446] Methods of determining the presence and severity of herpes-mediated encephalitis are well known in the art, and are described, for example, in Bonkowsky JL et al. (Herpes simplex virus central nervous system relapse during treatment of infantile spasms with corticotropin. Pediatrics. 2006 May; 117(5):e1045-8) and Khan OA, et al. (Herpes encephalitis presenting as mild aphasia: case report. BMC Fam Pract.2006 Mar 24; 7:22). Each method represents a separate embodiment of the present disclosure. [0447] In another embodiment, the present disclosure provides a method of treating or reducing the incidence of a disease, disorder, or symptom associated with an HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0448] In another embodiment, the disease, disorder, or symptom secondary to an HSV infection is oral lesions. In another embodiment, the disease, disorder, or symptom is genital lesions. In another embodiment, the disease, disorder, or symptom is oral ulcers. In another embodiment, the disease, disorder, or symptom is genital ulcers. In another embodiment, the disease, disorder, or symptom is fever. In another embodiment, the disease, disorder, or symptom is headache. In another embodiment, the disease, disorder, or symptom is muscle ache. In another embodiment, 295 11979815v1 P-627574-PC the disease, disorder, or symptom is swollen glands in the groin area. In another embodiment, the disease, disorder, or symptom is painful urination. In another embodiment, the disease, disorder, or symptom is vaginal discharge. In another embodiment, the disease, disorder, or symptom is blistering. In another embodiment, the disease, disorder, or symptom is flu-like malaise. In another embodiment, the disease, disorder, or symptom is keratitis. In another embodiment, the disease, disorder, or symptom is herpetic whitlow. In another embodiment, the disease, disorder, or symptom is Bell's palsy. In another embodiment, the disease, disorder, or symptom is herpetic erythema multiforme. In another embodiment, the disease, disorder, or symptom is a lower back symptom (e.g. numbness, tingling of the buttocks or the area around the anus, urinary retention, constipation, and impotence). In another embodiment, the disease, disorder, or symptom is a localized eczema herpeticum. In another embodiment, the disease, disorder, or symptom is a disseminated eczema herpeticum. In another embodiment, the disease, disorder, or symptom is a herpes gladiatorum. In another embodiment, the disease, disorder, or symptom is a herpetic sycosis. In another embodiment, the disease, disorder, or symptom is an esophageal symptom (e.g. difficulty swallowing or burning, squeezing throat pain while swallowing, weight loss, pain in or behind the upper chest while swallowing). In another embodiment, the disease, disorder, or symptom is any other disease, disorder, or symptom that is known in the art. Each disease, disorder, and symptom represents a separate embodiment of the present disclosure. [0449] Thus, in some embodiments, the compositions and methods of the instant disclosure treat, suppress, inhibit, or reduce the incidence of the infection itself, while in another embodiment, the compositions and methods of the instant disclosure treat, suppress, inhibit, or reduce the incidence of primary symptoms of the infection, while in another embodiment, the compositions and methods of the instant disclosure treat, suppress, inhibit, or reduce the incidence of secondary symptoms of the infection. It is to be understood that the compositions and methods of the instant disclosure may affect any combination of the infection, the primary symptoms caused by the infection, and secondary symptoms related to the infection. [0450] The HSV infection that is treated or ameliorated by methods and compositions of the present disclosure is, in another embodiment, a genital HSV infection. In another embodiment, the HSV infection is an oral HSV infection. In another embodiment, the HSV infection is an ocular HSV infection. In another embodiment, the HSV infection is a dermatologic HSV infection. In another embodiment, the HSV infection is a neurological HSV infection. In another embodiment, the HSV infection is a neuronal HSV infection. 296 11979815v1 P-627574-PC [0451] In another embodiment, the present disclosure provides a method of reducing the incidence of a disseminated HSV infection in a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0452] In another embodiment, the present disclosure provides a method of reducing the incidence of a neonatal HSV infection in an offspring of a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0453] In another embodiment, the present disclosure provides a method of reducing a transmission of an HSV infection from a subject to an offspring thereof, the method comprising the step of administering to said subject a composition of the present disclosure. [0454] In another embodiment, the offspring is an infant. In another embodiment, the transmission that is reduced or inhibited is transmission during birth. In another embodiment, transmission during breastfeeding is reduced or inhibited. In another embodiment, the transmission that is reduced or inhibited is any other type of parent-to-offspring transmission known in the art. [0455] In another embodiment, the present disclosure provides a method of reducing a severity of a neonatal HSV infection in an offspring of a subject, the method comprising the step of administering to said subject a composition of the present disclosure. [0456] In some embodiments, the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition comprising: (a) an RNA encoding HSV gC protein or fragment thereof; (b) an RNA encoding HSV gE protein or fragment thereof; and (c) an adjuvant. In another embodiment, the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence of an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition comprising: (a) an RNA encoding HSV gC protein or fragment thereof, wherein said fragment comprises either a C3b-binding domain thereof, a properdin interfering domain thereof, a C5 interfering domain thereof, or a fragment of said C3b-binding domain, properdin interfering domain, or C5-interfering domain; (b) an RNA encoding HSV gE protein or fragment thereof, wherein said fragment comprises AA 24-409, 24-405, or a fragment thereof; and (c) an adjuvant. [0457] In another embodiment, the present disclosure provides a method of treating an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of suppressing an HSV infection in a subject infected with HIV, the method 297 11979815v1 P-627574-PC comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of inhibiting an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of reducing the incidence of an HSV infection in a subject infected with HIV, the method comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of preventing an HIV infection, the method comprising the step of administering to said subject an HSV composition of the present disclosure. In some embodiments, HSV infection increases the risk of HIV infection, and protection against HSV infection decreases the risk of HIV infection. Thus, in some embodiments, the present disclosure provides a method of decreasing the risk of an HIV infection, the method comprising the step of administering to said subject a composition of the present disclosure. [0458] In some embodiments, the composition for use in the methods of the present disclosure elicits an immune response against HSV. In another embodiment, the composition for use in the methods of the present disclosure elicits an immune response against HSV-1. In another embodiment, the composition for use in the methods of the present disclosure elicits an immune response against HSV-2. In another embodiment, the composition comprises RNAs encoding gD and gC proteins, or immunogenic fragments thereof. In another embodiment, the composition comprises RNAs encoding gE and gD proteins, or immunogenic fragments thereof. In another embodiment, the composition comprises RNAs encoding gC and gE proteins, or immunogenic fragments thereof. In another embodiment, the composition comprises RNAs encoding gE, gD, and gC proteins, or immunogenic fragments thereof. In another embodiment, the composition comprises RNAs encoding gE, gD, or gC protein, or immunogenic fragments thereof. In another embodiment, the proteins encoded by the RNAs are HSV-1 proteins, or immunogenic fragments thereof. In another embodiment, the proteins encoded by the RNAs are HSV-2 proteins, or immunogenic fragments thereof. In another embodiment, the proteins encoded by the RNAs comprise both HSV-1 and HSV-2 proteins, or immunogenic fragments thereof. [0459] It is to be understood that, in some embodiments, a subject according to any of the embodiments described herein may be a subject infected with, or in another embodiment, susceptible to infection with HSV. In some embodiments, a subject may be infected with, or in another embodiment, susceptible to infection with at least one other pathogen. In some embodiments, a subject may be immunocompromised. In some embodiments, the subject is 298 11979815v1 P-627574-PC infected by HSV, while in another embodiment, the subject is at risk for infection by HSV, which in some embodiments, is a subject who is a neonate, in another embodiment, immunocompromised, in another embodiment, elderly, and in another embodiment, an immunocompromised neonate or an immunocompromised elderly subject. [0460] In another embodiment, the compositions of the present disclosure and their related uses may suppress, inhibit, prevent or treat an HIV infection in a subject. In some embodiments, the compositions of the present disclosure and their related uses may treat secondary complications of HIV infection, which in some embodiments, are opportunistic infections, neoplasms, neurologic abnormalities, or progressive immunologic deterioration. In another embodiment, the methods comprise treating acquired immunodeficiency syndrome (AIDS). In another embodiment, the methods comprise treating a decline in the number of CD4+ T lymphocytes. [0461] In another embodiment, the present disclosure provides a method of reducing HIV-1 transmission to an offspring, the method comprising the step of administering to a subject a composition of the present disclosure. As is known in the art, HSV-2 infection increases HIV-1 viral shedding in genital secretions (Nagot N et al., Reduction of HIV-1 RNA levels with therapy to suppress herpes simplex virus. N Engl J Med. 2007 Feb 22; 356(8):790-9). Thus, methods of the present disclosure of inhibiting HSV-2 infection are also efficacious for reducing HIV-1 transmission to an offspring. In another embodiment, the mutant HSV strain is an HSV-1 strain. In another embodiment, the mutant HSV strain is an HSV-2 strain. [0462] In another embodiment, the present disclosure provides a method of reducing HIV-1 transmission to a sexual partner, the method comprising the step of administering to a subject a composition of the present disclosure. As is known in the art, HSV-2 infection increases HIV-1 viral shedding in genital secretions. Thus, methods of the present disclosure of inhibiting HSV-2 infection are also efficacious for reducing HIV-1 transmission to a sexual partner. In another embodiment, the mutant HSV strain is an HSV-1 strain. In another embodiment, the mutant HSV strain is an HSV-2 strain. [0463] In another embodiment, the present disclosure provides a method of reducing susceptibility to HIV-1, the method comprising the step of administering to a subject a composition of the present disclosure. As is known in the art, HSV-2 infection increases HIV-1 replication (Ouedraogo A et al., Impact of suppressive herpes therapy on genital HIV-1 RNA among women taking antiretroviral therapy: a randomized controlled trial. AIDS. 2006 Nov 28;20(18):2305-13). Thus, methods of the present disclosure of inhibiting HSV-2 infection are also efficacious for reducing 299 11979815v1 P-627574-PC susceptibility to HIV-1. In another embodiment, the mutant HSV strain is an HSV-1 strain. In another embodiment, the mutant HSV strain is an HSV-2 strain. [0464] Thus, in some embodiments, the present disclosure provides a method of inhibiting a primary HSV infection in an HIV-infected subject, comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of treating or reducing the incidence of an HSV infection in an HIV-infected subject, comprising the step of administering to said subject a composition of the present disclosure. In another embodiment, the present disclosure provides a method of inhibiting a flare, recurrence, or HSV labialis following a primary HSV infection in an HIV-infected subject, the method comprising the step of administering to said subject a composition of the present disclosure. In some embodiments, administration of a composition of the present disclosure an anti-HSV immune response. [0465] In another embodiment, the present disclosure provides a method for inducing an immune response in a subject, the method comprising the step of administering to said subject a nucleoside RNA composition of the present disclosure. In another embodiment, the immune response comprises a CD4 immune response. In another embodiment, the immune response comprises a CD8 immune response. In another embodiment, the immune response comprises a T follicular helper cell immune response. In another embodiment, the immune response comprises a germinal center B cell immune response. In another embodiment, the immune response comprises an IgG antibody response to gC2, gD2, gE2, immunogenic fragments thereof, or a combination thereof. [0466] In another embodiment, the present disclosure provides a method of treating a Herpes Simplex Virus (HSV) infection in a subject, the method comprising the step of intramuscularly administering to said subject a nucleoside RNA composition of the present disclosure. In another embodiment, the disclosure provides a method of suppressing, inhibiting, or reducing the incidence of a Herpes Simplex Virus (HSV) infection in a subject, the method comprising the step of intramuscularly administering to said subject a nucleoside RNA composition of the present disclosure. [0467] In another embodiment, the present disclosure provides a method of treating, suppressing, inhibiting, or reducing the incidence a Herpes Simplex Virus (HSV) infection in a subject, the method comprising the step of administering to said subject a nucleoside RNA composition of the present disclosure and further comprising administering to said subject one or more polypeptide compositions comprising the proteins or immunogenic fragments thereof that are encoded by the 300 11979815v1 P-627574-PC nucleoside RNA as described herein. In one embodiment, the polypeptide composition comprises (a) a composition comprising HSV gD or an immunogenic fragment thereof, (b) a composition comprising HSV gC or an immunogenic fragment thereof, and (c) a composition comprising HSV gE or an immunogenic fragment thereof. [0468] In another embodiment, the disclosure provides a method of of a Herpes Simplex Virus (HSV) infection in a subject, the method comprising the step of intramuscularly administering to said subject a nucleoside RNA composition of the present disclosure. [0469] In another embodiment, the present disclosure provides a method as described herein wherein: the HSV-2 gC or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 28, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 241, or SEQ ID NO: 242; the HSV-2 gD or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 110, SEQ ID NO: 234, or SEQ ID NO: 120; and/or the HSV-2 gE or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 17 or SEQ ID NO: 18. [0470] In another embodiment, the present disclosure provides a method as described herein wherein: the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 25, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239, or SEQ ID NO: 246; the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 23, SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, or SEQ ID NO: 247; and/or the RNA encoding HSV-2 gE or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 27, SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, or SEQ ID NO: 148. 301 11979815v1 P-627574-PC [0471] In another embodiment, the present disclosure provides a method as described herein wherein: the HSV-2 gC or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 224; the HSV-2 gD or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 5; and/or the HSV-2 gE or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 17. [0472] In another embodiment, the present disclosure provides a method as described herein wherein: the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 246; the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 247; and/or the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 145. [0473] In another embodiment, the present disclosure provides a method as described herein wherein: the HSV-1 gC or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 240; the HSV-1 gD or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 134; and/or the HSV-1 gE or immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 248, or SEQ ID NO: 140. [0474] In another embodiment, the present disclosure provides a method as described herein wherein: the RNA encoding HSV-1 gC or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 24; the RNA encoding HSV-1 gD or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 22; and/or the RNA encoding HSV-1 gE or an immunogenic fragment thereof comprises a sequence at least 85% identical to SEQ ID NO: 26. 302 11979815v1 P-627574-PC [0475] In another embodiment, the present disclosure provides a method as described herein wherein the signal sequence is any signal sequence listed in Table 3. [0476] In one embodiment, the signal sequence is a viral signal sequence. [0477] In one embodiment, the viral signal sequence is an HSV signal sequence. [0478] In one embodiment, the HSV signal sequence is an HSV gD signal sequence. [0479] In one embodiment, the HSV gD signal sequence is an HSV-1 gD signal sequence. [0480] In one embodiment, the HSV-1 gD signal sequence encodes a polypeptide comprising a sequence at least 85% identical to SEQ ID NO: 34 or SEQ ID NO: 35. [0481] In one embodiment, wherein the HSV-1 gD signal sequence encodes a polypeptide comprising or consisting of a sequence at least 85% identical to SEQ ID NO: 35. [0482] In one embodiment, the HSV gD signal sequence is an HSV-2 gD signal sequence. [0483] In one embodiment, the HSV-2 gD signal sequence encodes a polypeptide comprising a sequence at least 85% identical to SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, or SEQ ID NO: 33. [0484] In one embodiment, the HSV-2 gD signal sequence encodes a polypeptide comprising or consisting of a sequence at least 85% identical to SEQ ID NO: 29 or SEQ ID NO: 31. [0485] In one embodiment, wherein the signal sequence is a codon optimized signal sequence. [0486] In one embodiment, the method comprises administering to said subject: a composition comprising RNA at least 85% identical to SEQ ID NO: 246; a composition comprising RNA at least 85% identical to SEQ ID NO: 247; and a composition comprising RNA at least 85% identical to SEQ ID NO: 145. Administration and Pharmaceutical Regimens [0487] Compositions of the present disclosure can be, in another embodiment, administered to a subject by any method known to a person skilled in the art, such as parenterally, paracancerally, transmucosally, transdermally, intramuscularly, intravenously, intra-dermally, subcutaneously, intra-peritonealy, intra-ventricularly, intra-cranially, intra-vaginally, intra-nasally, intra-tumorally, or topically. [0488] “Administering,” in another embodiment, refers to directly introducing into a subject by injection or other means a composition of the present disclosure. In another embodiment, “administering” refers to contacting a cell of the subject’s immune system with a composition or RNA encoding HSV protein or mixture thereof. 303 11979815v1 P-627574-PC [0489] In another embodiment of the methods and compositions of the present disclosure, the compositions are administered orally, and are thus formulated in a form suitable for oral administration, i.e. as a solid or a liquid preparation. Suitable solid oral formulations include tablets, capsules, pills, granules, pellets and the like. Suitable liquid oral formulations include solutions, suspensions, dispersions, emulsions, oils and the like. In another embodiment of the present disclosure, the active ingredient is formulated in a capsule. In accordance with this embodiment, the compositions of the present disclosure comprise, in addition to the active compound and the inert carrier or diluent, a hard gelating capsule. [0490] In other embodiments, the pharmaceutical compositions are administered by intravenous, intra-arterial, or intramuscular injection of a liquid preparation. Suitable liquid formulations include solutions, suspensions, dispersions, emulsions, oils and the like. In another embodiment, the pharmaceutical compositions are administered intravenously and are thus formulated in a form suitable for intravenous administration. In another embodiment, the pharmaceutical compositions are administered intra-arterially and are thus formulated in a form suitable for intra-arterial administration. In another embodiment, the pharmaceutical compositions are administered intramuscularly and are thus formulated in a form suitable for intramuscular administration. [0491] In another embodiment, the pharmaceutical compositions are administered topically to body surfaces and are thus formulated in a form suitable for topical administration. Suitable topical formulations include gels, ointments, creams, lotions, drops and the like. For topical administration, the compositions or their physiologically tolerated derivatives are prepared and applied as solutions, suspensions, or emulsions in a physiologically acceptable diluent with or without a pharmaceutical carrier. [0492] In another embodiment, the composition is administered as a suppository, for example a rectal suppository or a urethral suppository. In another embodiment, the pharmaceutical composition is administered by subcutaneous implantation of a pellet. In another embodiment, the pellet provides for controlled release of agent over a period of time. [0493] In a preferred embodiment, pharmaceutical compositions are administered intramuscularly, subcutaneously or intradermally. [0494] “Effective dosage” of the RNA, refers, in another embodiment, to an amount sufficient to exert a therapeutic effect. In another embodiment, the term refers to an amount sufficient to elicit expression of a detectable amount of the encoded protein. Each possibility represents a separate embodiment of the present disclosure. 304 11979815v1 P-627574-PC [0495] Methods for measuring the dose of an RNA encoding an HSV glycoprotein, or an immunogenic fragment thereof (e.g. in human subjects) are well known in the art, and include, for example, dose-escalating trials. Each method represents a separate embodiment of the present disclosure. [0496] In some embodiments, any of the HSV compositions of and for use in the methods of this disclosure will comprise an RNA encoding HSV protein or an immunogenic fragment thereof or combination of RNAs encoding HSV proteins or immunogenic fragments thereof, of the present disclosure, in any form or embodiment as described herein. In some embodiments, any of the compositions of and for use in the methods will consist of an RNA encoding an HSV protein, or immunogenic fragment thereof, or combination of RNA encoding HSV proteins, or immunogenic fragments thereof, of the present disclosure, in any form or embodiment as described herein. In some embodiments, the compositions of this disclosure will consist essentially of an RNA encoding an HSV protein, or immunogenic fragment thereof, or combination of RNAs encoding HSV proteins, or an immunogenic fragments thereof, of the present disclosure, in any form or embodiment as described herein. In some embodiments, the term “comprise” refers to the inclusion of RNA encoding other HSV proteins, or immunogenic fragments thereof, as well as inclusion of RNA encoding other proteins that may be known in the art. In some embodiments, the term “consisting essentially of” refers to a composition, which has the RNA encoding a specific HSV protein or fragment thereof. However, other components may be included that are not involved directly in the utility of the RNA(s) encoding HSV protein(s). In some embodiments, the term “consisting” refers to a composition having an RNA encoding particular HSV protein or fragment or combination of RNAs encoding HSV proteins or fragments of the present disclosure, in any form or embodiment as described herein. [0497] In another embodiment, the present disclosure provides a composition for treating HSV-1 or a symptom or manifestation thereof, the composition comprising an RNA of the present disclosure. [0498] In another embodiment, the present disclosure provides a composition for treating HSV-2 or a symptom or manifestation thereof, the composition comprising an RNA of the present disclosure. [0499] It is to be understood that compositions, and methods of the present disclosure may be used in non-HSV herpesvirus as well, which in some embodiments, proteins gD, gE, or gC proteins that are, in some embodiments, 70% homologous, in another embodiment, 80% homologous, in 305 11979815v1 P-627574-PC another embodiment, 85% homologous, in another embodiment, 90% homologous, in another embodiment, 95% homologous, in another embodiment, 98% homologous, and in another embodiment, 100% homologous to the gD, gE, or gC proteins of HSV-1, or in another embodiment, of HSV-2. In some embodiments, such compositions may be useful in suppressing, inhibiting, preventing, or treating, cancers, or in another embodiment, tumors. In some embodiments, non-HSV herpesvirus comprise Varicella Zoster Virus (VZV), Epstein-Barr virus (EBV), EBNA, cytomegalovirus (CMV), and human herpesvirus-6 (HHV-6). [0500] In another embodiment, of methods of the present disclosure, a composition of the present disclosure is administered once. In another embodiment, the composition is administered twice. In another embodiment, the composition is administered three times. In another embodiment, the composition is administered four times. In another embodiment, the composition is administered at least four times. In another embodiment, the composition is administered more than four times. [0501] In another embodiment, the dosage is a daily dose. In another embodiment, the dosage is a weekly dose. In another embodiment, the dosage is a monthly dose. In another embodiment, the dosage is an annual dose. In another embodiment, the dose is one is a series of a defined number of doses. In another embodiment, the dose is a one-time dose. [0502] In some embodiments, any of the booster doses described hereinabove is administered following a priming dose comprising one or modified more RNAs encoding HSV-1 proteins or immunogenic fragments thereof. In another embodiment, any of the booster doses described hereinabove is administered following a priming vaccination comprising one or more modified more RNAs encoding HSV-2 proteins or immunogenic fragments thereof. [0503] In some embodiments, a subject is immunized with a single administration of the composition. In another embodiment, a subject is immunized with a single dose. In another embodiment, a subject is immunized with two doses. In another embodiment, a subject is immunized with three doses. In another embodiment, a subject is immunized with four doses. In another embodiment, a subject is immunized with five doses. [0504] In some embodiments, all the components of the composition are provided in equal concentrations. According to this aspect and in some embodiments, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are provided in a ratio of 1:1:1. In another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are provided in a ratio of 5:2:5. In another embodiment, RNAs encoding gC and gD, or immunogenic fragments thereof, are provided in a ratio of 1:1. In another embodiment, RNAs encoding gC and gE, or immunogenic 306 11979815v1 P-627574-PC fragments thereof, are provided in a ratio of 1:1. In another embodiment, RNAs encoding gD and gE, or immunogenic fragments thereof, are provided in a ratio of 1:1. [0505] In some embodiments, RNAs encoding gC, gD, gE, immunogenic fragments thereof, or a combination thereof, or combined with other HSV glycoproteins, are administered in a single composition at the same site and by the same route, while in another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered in separate compositions at separate sites but by the same route of administration, or in another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered in separate compositions at separate sites and by different routes of administration, or in another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered in separate compositions at the same site and by different routes of administration (e.g. injection and topical). [0506] In some embodiments, the methods of the present disclosure include a one-time or single administration of compositions comprising one or more nucleoside RNAs of the present disclosure. In another embodiment, the methods of the present disclosure include administration of compositions comprising one or more nucleoside RNAs in a prime and boost approach. In some embodiments, the methods of the present disclosure further comprise the step of administering to said subject one or more additional administrations of said nucleoside RNA composition subsequent to the first administration. [0507] In another embodiment, the methods of the present disclosure comprise administering a composition comprising one or more nucleoside RNAs encoding one or more HSV glycoproteins, or immunogenic fragments thereof, as a first administration and a composition comprising one or more HSV glycoproteins, or immunogenic fragments thereof, as a second or subsequent administration. In some embodiments, the HSV glycoproteins, or immunogenic fragments thereof, encoded by an RNA in the first (or prime) administration are the same glycoproteins, or fragments thereof, in the second or subsequent (or boost) administration. In another embodiment, a composition comprising one or more HSV glycoproteins, or immunogenic fragments thereof, is administered as a first administration, and a composition comprising one or more nucleoside RNAs encoding one or more HSV glycoproteins, or immunogenic fragments thereof, is administered as a second or subsequent administration. Each possibility represents a separate embodiment of the present disclosure. [0508] In another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered simultaneously followed by a booster dose of RNA encoding gD without 307 11979815v1 P-627574-PC RNAs encoding gC or gE, or immunogenic fragments thereof. In another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered simultaneously followed by a booster dose of RNA encoding gC without RNAs encoding gD or gE, or immunogenic fragments thereof,. In another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered simultaneously followed by a booster dose of RNA encoding gE without RNAs encoding gD or gC, or immunogenic fragments thereof,. In another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered simultaneously followed by a booster dose of RNAs encoding gC and gD, or immunogenic fragments thereof, without RNAs encoding gE, or immunogenic fragments thereof,. In another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered simultaneously followed by a booster dose of RNAs encoding gC and gE, or immunogenic fragments thereof, without RNAs encoding gD, or immunogenic fragments thereof,. In another embodiment, RNAs encoding gC, gD, and gE, or immunogenic fragments thereof, are administered simultaneously followed by a booster dose of RNAs encoding gD and gE, or immunogenic fragments thereof, without RNAs encoding gE, or immunogenic fragments thereof,. In one embodiment the booster administration is performed at the same site and by the same mode of administration as the priming administration. In another embodiment, the booster administration is performed at a different site from the priming administration but by the same mode of administration as the priming administration. In one embodiment the booster administration is performed at the same site but by different mode of administration than priming administration. In another embodiment, the booster administration is performed at a different site and by different mode of administration than priming administration. [0509] In some embodiments, the modified RNA induces a detectably lower innate immune response than the same quantity of unmodified RNA having the same sequence. [0510] In some embodiments, the effectiveness of the compositions and methods of the present disclosure are dependent on the presence of complement, while in another embodiment, the compositions and methods of the present disclosure are not dependent on the presence of complement. In some embodiments, the effectiveness of some of the compositions for use in the methods of the present disclosure are dependent on the presence of complement, while others are not. In some embodiments, the anti-gC antibody is dependent on complement for its effectiveness against HSV. 308 11979815v1 P-627574-PC [0511] In some embodiments, complement is an important contributor to innate and acquired immunity. In some embodiments, complement activation facilitates virus neutralization by particle phagocytosis and lysis, functions as a chemoattractant for neutrophils and macrophages, and enhances B and T cell responses. In some embodiments, HSV-1 gC, or an immunogenic fragment thereof, binds complement C3b and blocks C5 and properdin interaction with C3b, which inhibit complement activation and complement-mediated virus neutralization. In some embodiments, a gC1 domain, or an immunogenic fragment thereof, that interacts with complement is located within amino acids 33 to 133 and blocks C5 and properdin binding to C3b, and In some embodiments, a gC1, or an immunogenic fragment thereof, domain that interacts with complement extends from amino acids 124 to 366 and directly binds C3b. In some embodiments, an HSV-1 gC mutant virus deleted in the C3b binding domain is more susceptible to complement-mediated virus neutralization in vitro and less virulent than wild-type (WT) virus in the mouse flank model. Therefore, in some embodiments, the interaction between gC1 and C3b enhances HSV-1 virulence, and in some embodiments, blocking the gC1 domain is effective in preventing or treating HSV-1 infection. [0512] In some embodiments, the compositions and methods of the present disclosure are for use in human subjects, while in another embodiment, they are for use in animal subjects. In another embodiment, the subject is mammalian. In another embodiment, the subject is any organism susceptible to infection by HSV. In some embodiments, the subject is murine, bovine, ovine, canine, feline, equine, porcine, etc. In some embodiments, the compositions and methods of the present disclosure are effective in male subjects. In another embodiment, the compositions and methods of the present disclosure are effective in female subjects. In some embodiments, the compositions and methods of the present disclosure are effective in seronegative subjects. In another embodiment, the compositions and methods of the present disclosure are effective in seropositive subjects. Pharmaceutical Formulations [0513] In some embodiments, a method of present disclosure further comprises mixing an RNA with a transfection reagent prior to the step of contacting. In another embodiment, a method of present disclosure further comprises administering an RNA together with a transfection reagent. In another embodiment, the transfection reagent is a cationic lipid reagent. For example, the RNA can be one or more RNAs comprising a nucleotide sequence at least 80%, at least 85%, at least 309 11979815v1 P-627574-PC 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6. [0514] In another embodiment, a transfection reagent is a lipid-based transfection reagent. In another embodiment, a transfection reagent is a protein-based transfection reagent. In another embodiment, a transfection reagent is a polyethyleneimine based transfection reagent. In another embodiment, a transfection reagent is calcium phosphate. In another embodiment, a transfection reagent is Lipofectin® or Lipofectamine®. In another embodiment, a transfection reagent is any other transfection reagent known in the art. [0515] In another embodiment, a transfection reagent forms a liposome. Liposomes, in another embodiment, increase intracellular stability, increase uptake efficiency and improve biological activity. [0516] In another embodiment, liposomes are hollow spherical vesicles composed of lipids arranged in a similar fashion as those lipids which make up the cell membrane. They have, in another embodiment, an internal aqueous space for entrapping water soluble compounds and range in size from 0.05 to several microns in diameter. In another embodiment, liposomes can deliver RNA to cells in a biologically active form (see Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; see generally ibid). [0517] Each type of transfection reagent represents a separate embodiment of the present disclosure. [0518] In another embodiment, an RNA of the present disclosure is encapsulated in a nanoparticle. Methods for nanoparticle packaging are well known in the art, and are described, for example, in Bose S, et al., (Role of Nucleolin in Human Parainfluenza Virus Type 3 Infection of Human Lung Epithelial Cells. J. Virol. 78:8146. 2004); Dong Y et al., Poly(d,l-lactide-co- glycolide)/montmorillonite nanoparticles for oral delivery of anticancer drugs. Biomaterials 26:6068. 2005); Lobenberg R. et al., (Improved body distribution of 14C-labelled AZT bound to nanoparticles in rats determined by radioluminography. J Drug Target 5:171.1998); Sakuma S R et al., (Mucoadhesion of polystyrene nanoparticles having surface hydrophilic polymeric chains in the gastrointestinal tract. Int J Pharm 177:161.1999); Virovic L et al., Novel delivery methods for treatment of viral hepatitis: an update. Expert Opin Drug Deliv 2:707.2005); and Zimmermann E et al., Electrolyte- and pH-stabilities of aqueous solid lipid nanoparticle (SLN) dispersions in 310 11979815v1 P-627574-PC artificial gastrointestinal media. Eur J Pharm Biopharm 52:203.2001). Each method represents a separate embodiment of the disclosure. [0519] In some embodiments, ψRNA is encapsulated in nanoparticles to improve efficiency of delivery and expression of ψRNA. Nanoparticle packaging involves condensing and encapsulating RNA into particles that are smaller than the pore of the nuclear membrane, using chemicals including poly-L-lysine and polyethylene glycol. In some embodiments, RNA is packaged into one of four nanoparticle formulations (PEI, PLL, PAE, and CK30PEG10k). Lipid Nanoparticles [0520] In some embodiments, nanoparticles used in the compositions and methods of the present disclosure comprise lipid nanoparticles as described in Cullis, P., & Hope, M. (n.d.). Lipid Nanoparticle Systems for Enabling Gene Therapies. Molecular therapy, 25(7), which is incorporated by reference herein in its entirety. [0521] In some embodiments, delivery of RNA (e.g., nucleoside modified RNA) comprises any suitable delivery method, including exemplary RNA transfection methods described elsewhere herein. In certain embodiments, delivery of a nucleoside-modified RNA to a subject comprises mixing the nucleoside-modified RNA with a transfection reagent prior to the step of contacting. In another embodiment, a method of present disclosure further comprises administering nucleoside-modified RNA together with the transfection reagent. In another embodiment, the transfection reagent is a cationic lipid reagent. For example, said RNAs can be one or more RNAs comprising a nucleotide sequence at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a sequence listed in Table 6. [0522] In another embodiment, the transfection reagent is a lipid-based transfection reagent. In another embodiment, the transfection reagent is a protein-based transfection reagent. In another embodiment, the transfection reagent is a polyethyleneimine based transfection reagent. In another embodiment, the transfection reagent is calcium phosphate. In another embodiment, the transfection reagent is Lipofectin®, Lipofectamine®, or TransIT®. In another embodiment, the transfection reagent is any other transfection reagent known in the art. [0523] In another embodiment, a transfection reagent forms a liposome. [0524] Liposomes, in another embodiment, increase intracellular stability, increase uptake efficiency and improve biological activity. In another embodiment, liposomes are hollow spherical vesicles composed of lipids arranged in a similar fashion as those lipids which make up the cell 311 11979815v1 P-627574-PC membrane. They have, in another embodiment, an internal aqueous space for entrapping water- soluble compounds and range in size from 0.05 to several microns in diameter. In another embodiment, liposomes can deliver RNA to cells in a biologically active form. [0525] In some embodiments, the composition comprises a lipid nanoparticle (LNP) and one or more nucleic acid molecules described herein. For example, In some embodiments, the composition comprises an LNP and one or more nucleoside-modified RNA molecules encoding one or more antigens, adjuvants, or a combination thereof. [0526] The term "lipid nanoparticle" refers to a particle having at least one dimension on the order of nanometers (e.g., 1-1,000 nm) which includes one or more lipids, for example a lipid of Formula (I), (II) or (III), as described in WO2016176330A1, which is incorporated by reference herein in its entirety. [0527] In some embodiments, lipid nanoparticles are included in a formulation comprising a nucleoside-modified RNA as described herein. In some embodiments, such lipid nanoparticles comprise a cationic lipid and one or more excipient selected from neutral lipids, charged lipids, steroids and polymer conjugated lipids (e.g., a pegylated lipid such as a pegylated lipid of structure (IV), such as compound IVa). In some embodiments, the nucleoside-modified RNA is encapsulated in the lipid portion of the lipid nanoparticle or an aqueous space enveloped by some or all of the lipid portion of the lipid nanoparticle, thereby protecting it from enzymatic degradation or other undesirable effects induced by the mechanisms of the host organism or cells e.g. an adverse immune response. [0528] In various embodiments, the lipid nanoparticles have a mean diameter of from about 30 nm to about 150 nm, from about 40 nm to about 150 nm, from about 50 nm to about 150 nm, from about 60 nm to about 130 nm, from about 70 nm to about 110 nm, from about 70 nm to about 100 nm, from about 80 nm to about 100 nm, from about 90 nm to about 100 nm, from about 70 to about 90 nm, from about 80 nm to about 90 nm, from about 70 nm to about 80 nm, or about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, or 150 nm, and are substantially non-toxic. In certain embodiments, the nucleoside-modified RNA, when present in the lipid nanoparticles, is resistant in aqueous solution to degradation with a nuclease. [0529] The LNP may comprise any lipid capable of forming a particle to which the one or more nucleic acid molecules are attached, or in which the one or more nucleic acid molecules are 312 11979815v1 P-627574-PC encapsulated. The term "lipid" refers to a group of organic compounds that are derivatives of fatty acids (e.g., esters) and are generally characterized by being insoluble in water but soluble in many organic solvents. Lipids are usually divided in at least three classes: (1) "simple lipids" which include fats and oils as well as waxes; (2) "compound lipids" which include phospholipids and glycolipids; and (3) "derived lipids" such as steroids. [0530] In some embodiments, the LNP comprises one or more cationic lipids, and one or more stabilizing lipids. Stabilizing lipids include neutral lipids and pegylated lipids. [0531] In some embodiments, the LNP comprises a cationic lipid. As used herein, the term "cationic lipid" refers to a lipid that is cationic or becomes cationic (protonated) as the pH is lowered below the pK of the ionizable group of the lipid, but is progressively more neutral at higher pH values. At pH values below the pK, the lipid is then able to associate with negatively charged nucleic acids. In certain embodiments, the cationic lipid comprises a zwitterionic lipid that assumes a positive charge on pH decrease. [0532] In certain embodiments, the cationic lipid comprises any of a number of lipid species which carry a net positive charge at a selective pH, such as physiological pH. Such lipids include, but are not limited to, N,N-dioleyl-N,N-dimethylammonium chloride (DODAC); N-(2,3- dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA); N,N-distearyl-N,N- dimethylammonium bromide (DDAB); N-(2,3- dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP); 3-(N— (Ν',Ν'- dimethylaminoethane)-carbamoyl)cholesterol (DC-Choi), N- (l-(2,3-dioleoyloxy)propyl)- N-2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoracetate (DOSPA), dioctadecylamidoglycyl carboxy spermine (DOGS), l,2-dioleoyl-3- dimethylammonium propane (DODAP), N,N-dimethyl-2,3-dioleoyloxy)propylamine (DODMA), and N-(l,2- dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide (DMRIE). Additionally, a number of commercial preparations of cationic lipids are available which can be used in the present disclosure. These include, for example, LIPOFECTIN® (commercially available cationic liposomes comprising DOTMA and l,2-dioleoyl-sn-3- phosphoethanolamine (DOPE), from GIBCO/BRL, Grand Island, N.Y.); LIPOFECTAMINE® (commercially available cationic liposomes comprising N-(l-(2,3- dioleyloxy)propyl)-N-(2- (sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoroacetate (DOSPA) and (DOPE), from GIBCO/BRL); and TRANSFECTAM® (commercially available cationic lipids comprising dioctadecylamidoglycyl carboxyspermine (DOGS) in ethanol from Promega Corp., Madison, Wis.). The following lipids are cationic and have a positive charge at below physiological pH: 313 11979815v1 P-627574-PC DODAP, DODMA, DMDMA, l,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1 ,2- dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA). [0533] In some embodiments, the cationic lipid is an amino lipid. Suitable amino lipids useful in the disclosure include those described in WO 2012/016184, incorporated herein by reference in its entirety. Representative amino lipids include, but are not limited to, l,2-dilinoleyoxy-3- (dimethylamino)acetoxypropane (DLin-DAC), l,2-dilinoleyoxy-3- morpholinopropane (DLin- MA), l,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), l,2-dilinoleylthio-3- dimethylaminopropane (DLin-S-DMA), l-linoleoyl-2-linoleyloxy-3- dimethylaminopropane (DLin-2-DMAP), l,2-dilinoleyloxy-3-trimethylaminopropane chloride salt (DLin-TMA.Cl), l,2- dilinoleoyl-3-trimethylaminopropane chloride salt (DLin-TAP.Cl), l,2-dilinoleyloxy-3-(N- methylpiperazino)propane (DLin-MPZ), 3-(N,N- dilinoleylamino)-l,2-propanediol (DLinAP), 3- (N,N-dioleylamino)-l,2-propanediol (DOAP), l,2-dilinoleyloxo-3-(2-N,N- dimethylamino)ethoxypropane (DLin-EG-DMA), and 2,2-dilinoleyl-4-dimethylaminomethyl- [l,3]-dioxolane (DLin-K-DMA). [0534] In certain embodiments, the cationic lipid is present in the LNP in an amount from about 30 to about 95 mole percent. In some embodiments, the cationic lipid is present in the LNP in an amount from about 30 to about 70 mole percent. In some embodiments, the cationic lipid is present in the LNP in an amount from about 40 to about 60 mole percent. In some embodiments, the cationic lipid is present in the LNP in an amount of about 50 mole percent. In some embodiments, the LNP comprises only cationic lipids. In certain embodiments, the LNP comprises one or more additional lipids which stabilize the formation of particles during their formation. [0535] Suitable stabilizing lipids include neutral lipids and anionic lipids. [0536] The term "neutral lipid" refers to any one of a number of lipid species that exist in either an uncharged or neutral zwitterionic form at physiological pH. [0537] Representative neutral lipids include diacylphosphatidylcholines, diacylphosphatidylethanolamines, ceramides, sphingomyelins, dihydro sphingomyelins, cephalins, and cerebrosides. [0538] Exemplary neutral lipids include, for example, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl- phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE) and dioleoyl-phosphatidylethanolamine 4-(N- 314 11979815v1 P-627574-PC maleimidomethyl)-cyclohexane-l- carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl- phosphatidylethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, l- stearioyl-2-oleoyl- phosphatidyethanol amine (SOPE), and l,2-dielaidoyl-sn-glycero-3- phophoethanolamine (transDOPE). In some embodiments, the neutral lipid is l,2-distearoyl-sn- glycero-3- phosphocholine (DSPC). [0539] In some embodiments, the LNPs comprise a neutral lipid selected from DSPC, DPPC, DMPC, DOPC, POPC, DOPE and SM. In various embodiments, the molar ratio of the cationic lipid (e.g., lipid of Formula (I)) to the neutral lipid ranges from about 2 : 1 to about 8: 1. [0540] In various embodiments, the LNPs further comprise a steroid or steroid analogue. [0541] In certain embodiments, the steroid or steroid analogue is cholesterol. In some of these embodiments, the molar ratio of the cationic lipid (e.g., lipid of Formula (I)) to cholesterol ranges from about 2:1 to 1:1. [0542] The term "anionic lipid" refers to any lipid that is negatively charged at physiological pH. These lipids include phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N- dodecanoylphosphatidylethanolamines, N- succinylphosphatidylethanolamines, N- glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids. [0543] In certain embodiments, the LNP comprises glycolipids (e.g., monosialoganglioside GMi). In certain embodiments, the LNP comprises a sterol, such as cholesterol. [0544] In some embodiments, the LNPs comprise a polymer conjugated lipid. The term "polymer conjugated lipid" refers to a molecule comprising both a lipid portion and a polymer portion. An example of a polymer conjugated lipid is a pegylated lipid. The term "pegylated lipid" refers to a molecule comprising both a lipid portion and a polyethylene glycol portion. Pegylated lipids are known in the art and include l-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG- s- DMG) and the like. [0545] In certain embodiments, the LNP comprises an additional, stabilizing - lipid which is a polyethylene glycol-lipid (pegylated lipid). Suitable polyethylene glycol- lipids include PEG- modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramides (e.g., PEG-CerC14 or PEG-CerC20), PEG-modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols. 315 11979815v1 P-627574-PC [0546] Representative polyethylene glycol-lipids include PEG-c-DOMG, PEG-c-DMA, and PEG- s-DMG. In some embodiments, the polyethylene glycol-lipid is N-[(methoxy poly(ethylene glycol)2ooo)carbamyl]-l,2-dimyristyloxlpropyl-3-amine (PEG-c-DMA). In some embodiments, the polyethylene glycol-lipid is PEG-c-DOMG). In other embodiments, the LNPs comprise a pegylated diacylglycerol (PEG-DAG) such as l-(monomethoxy-polyethyleneglycol)-2,3- dimyristoylglycerol (PEG-DMG), a pegylated phosphatidylethanoloamine (PEG-PE), a PEG succinate diacylglycerol (PEG-S-DAG) such as 4-0-(2',3'-di(tetradecanoyloxy)propyl-l-0-(co- methoxy(polyethoxy)ethyl)butanedioate (PEG-S-DMG), a pegylated ceramide (PEG- cer), or a PEG dialkoxypropylcarbamate such as Q-methoxy(polyethoxy)ethyl-N-(2,3- di(tetradecanoxy)propyl)carbamate or 2,3-di(tetradecanoxy)propyl-N-(co- methoxy(polyethoxy)ethyl)carbamate. In various embodiments, the molar ratio of the cationic lipid to the pegylated lipid ranges from about 100: 1 to about 25: 1. [0547] In certain embodiments, the additional lipid is present in the LNP in an amount from about 1 to about 10 mole percent. In some embodiments, the additional lipid is present in the LNP in an amount from about 1 to about 5 mole percent. In some embodiments, the additional lipid is present in the LNP in about 1 mole percent or about 1.5 mole percent. [0548] In certain embodiments, the LNP comprises one or more targeting moieties which are capable of targeting the LNP to a cell or cell population. For example, In some embodiments, the targeting moiety is a ligand which directs the LNP to a receptor found on a cell surface. [0549] In certain embodiments, the LNP comprises one or more internalization domains. For example, in some embodiments, the LNP comprises one or more domains which bind to a cell to induce the internalization of the LNP. For example, in some embodiments, the one or more internalization domains bind to a receptor found on a cell surface to induce receptor-mediated uptake of the LNP. In certain embodiments, the LNP is capable of binding a biomolecule in vivo, where the LNP-bound biomolecule can then be recognized by a cell-surface receptor to induce internalization. For example, in some embodiments, the LNP binds systemic ApoE, which leads to the uptake of the LNP and associated cargo. [0550] Other exemplary LNPs and their manufacture are described in the art, for example in WO2016176330A1, U.S. Patent Application Publication No. US20120276209, Semple et al., 2010, Nat Biotechnol., 28(2): 172-176; Akinc et al., 2010, Mol Ther., 18(7): 1357-1364; Basha et al., 2011, Mol Ther, 19(12): 2186-2200; Leung et al., 2012, J Phys Chem C Nanomater Interfaces, 116(34): 18440-18450; Lee et al., 2012, Int J Cancer., 131(5): E781-90; Belliveau et al., 2012, 316 11979815v1 P-627574-PC Mol Ther nucleic Acids, 1 : e37; Jayaraman et al., 2012, Angew Chem Int Ed Engl., 51(34): 8529- 8533; Mui et al., 2013, Mol Ther Nucleic Acids.2, el39; Maier et al., 2013, Mol Ther., 21(8): 1570- 1578; and Tarn et al., 2013, Nanomedicine, 9(5): 665-74, each of which are incorporated by reference in their entirety. [0551] In another embodiment, methods of the present disclosure comprise administering an RNAs encoding one or more HSV glycoproteins, or immunogenic fragments thereof and a pharmaceutically acceptable carrier or diluent. In other embodiments, pharmaceutically acceptable carriers for liquid formulations may be aqueous or non-aqueous solutions, suspensions, emulsions or oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Examples of oils are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, and fish-liver oil. [0552] As used herein “pharmaceutically acceptable carriers or diluents” are well known to those skilled in the art. [0553] In another embodiment, the pharmaceutical compositions provided herein are controlled- release compositions, i.e. compositions in which the compound is released over a period of time after administration. Controlled- or sustained-release compositions include formulation in lipophilic depots (e.g., fatty acids, waxes, oils). In another embodiment, the composition is an immediate-release composition, i.e. a composition in which the entire compound is released immediately after administration. [0554] Each of the additives, excipients, formulations and methods of administration represents a separate embodiment of the present disclosure. [0555] In another embodiment, the present disclosure provides a kit comprising a reagent utilized in performing a method of the present disclosure. In another embodiment, the present disclosure provides a kit comprising a composition, tool, or instrument of the present disclosure. Certain Embodiments 1. A combination comprising: (a) a first composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and 317 11979815v1 P-627574-PC (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, for use in a method of inhibiting, suppressing, or reducing the incidence of an HSV oral mucosal infection, an HSV encephalitis infection, or any combination thereof, in a subject. 2. A combination comprising: (a) a first composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, for use in a method of treating an HSV oral mucosal infection, an HSV encephalitis infection, or any combination thereof, in a subject. 3. The combination of embodiment 1 or embodiment 2, wherein said infection is a primary infection or a secondary infection. 4. A combination comprising: (a) a first composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, for use in a method of preventing establishment of a latent HSV infection in a subject. 5. A combination comprising: (a) a first composition comprising RNA encoding Herpes Simplex Virus (HSV) glycoprotein D (gD) or an immunogenic fragment thereof, (a) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (b) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, 318 11979815v1 P-627574-PC for use in a method of decreasing, suppressing, or inhibiting an HSV infection of the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or any combination thereof in a subject. 6. The combination of any one of embodiments 1-5, wherein one or more of said RNAs comprise one or more ribonucleic acids that are nucleoside modified. 7. The combination of embodiment 6, wherein the one or more nucleoside modified ribonucleic acids comprise one or more m5D (5-methyldihydrouridine) residues. 8. The combination of embodiment 7, wherein the one or more nucleoside modified ribonucleic acids comprise one or more pseudouridine residues. 9. The combination of embodiment 8, wherein said one or more pseudouridine residues comprise m1Ψ (1-methylpseudouridine). 10. The combination of embodiment 8 or 9, wherein said one or more pseudouridine residues comprise m1acp3Ψ (1-methyl-3-(3-amino-5-carboxypropyl)pseudouridine, Ψm (2′-O- methylpseudouridine, m5D (5-methyldihydrouridine), m3Ψ (3-methylpseudouridine), or any combination thereof. 11. The combination of any one of embodiments 1-10, wherein the HSV gC or an immunogenic fragment thereof is an HSV-2 gC or immunogenic fragment thereof, the HSV gD or an immunogenic fragment thereof is an HSV-2 gD or immunogenic fragment thereof, the HSV gE or an immunogenic fragment thereof is an HSV-2 gE or immunogenic fragment thereof, or any combination thereof. 12. The combination of embodiment 11, wherein: the HSV-2 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 28, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 241, or SEQ ID NO: 242; the HSV-2 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 110, SEQ ID NO: 234, or SEQ ID NO: 120; and/or the HSV-2 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 17 or SEQ ID NO: 18. 13. The combination of embodiment 11 or 12, wherein: the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 25, SEQ ID NO: 119, SEQ ID 319 11979815v1 P-627574-PC NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239, or SEQ ID NO: 246; the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 23, SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, or SEQ ID NO: 247; the RNA encoding HSV-2 gE or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 27, SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, or SEQ ID NO: 148, or any combination thereof. 14. The combination of any one of embodiments 11-13, wherein: the HSV-2 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 224; the HSV-2 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 5; and/or the HSV-2 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 17. 15. The combination of any one of embodiments 11–14, wherein the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 246; the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 247; and/or the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 145. 16. The method of claim 15, wherein: (a) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 246; (b) the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 247; and (c) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 145. 320 11979815v1 P-627574-PC 17. The combination of any one of embodiments 1-10, wherein the HSV gC or an immunogenic fragment thereof is an HSV-1 gC or immunogenic fragment thereof, the HSV gD or an immunogenic fragment thereof is an HSV-1 gD or immunogenic fragment thereof, and/or the HSV gE or an immunogenic fragment thereof is an HSV-1 gE or immunogenic fragment thereof. 18. The combination of embodiment 17, wherein: the HSV-1 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 240; the HSV-1 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 134; and/or the HSV-1 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 248, or SEQ ID NO: 140. 19. The combination of embodiment 17 or 18, wherein: the RNA encoding HSV-1 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 24; the RNA encoding HSV-1 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 22; and/or the RNA encoding HSV-1 gE or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 26. 20. The combination of any one of embodiments 1-19, wherein the immunogenic fragment of HSV gC comprises the ectodomain of HSV gC, the immunogenic fragment of HSV gD comprises the ectodomain of HSV gD, and/or the immunogenic fragment of HSV gE comprises the ectodomain of HSV gE. 21. The combination of any one of embodiments 1-20, wherein said composition further comprises one or more RNAs encoding a) HSV glycoprotein B (gB) or immunogenic fragment thereof, b) HSV glycoprotein H (gH) or immunogenic fragment thereof, c) HSV glycoprotein L (gL) or immunogenic fragment thereof, d) HSV glycoprotein I (gI) or immunogenic fragment thereof, or e) any combination thereof. 22. The combination of any one of embodiments 1-21, wherein one or more of said RNAs further comprise a signal sequence encoding a signal peptide. 321 11979815v1 P-627574-PC 23. The combination of embodiment 22, wherein the signal sequence is or comprises any signal sequence listed in Table 4. 24. The combination of embodiment 23, wherein the signal peptide is an IL-2 signal peptide. 25. The combination of embodiment 23 or 24, wherein the signal peptide is a viral signal peptide. 26. The combination of embodiment 24, wherein the viral signal peptide is an HSV signal peptide. 27. The combination of embodiment 26, wherein the HSV signal peptide is an HSV gD signal peptide. 28. The combination of embodiment 27, wherein the HSV gD signal peptide is an HSV-1 gD signal peptide. 29. The combination of embodiment 28, wherein the HSV-1 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 34 or SEQ ID NO: 35. 30. The combination of embodiment 28 or 29, wherein the HSV-1 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 35. 31. The combination of embodiment 27, wherein the HSV gD signal peptide is an HSV-2 gD signal sequence. 32. The combination of embodiment 31, wherein the HSV-2 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, or SEQ ID NO: 33. 33. The combination of embodiment 31 or 32, wherein the HSV-2 gD signal peptides comprises an amino acid sequence at least 85% identical to SEQ ID NO: 29 or SEQ ID NO: 31. 34. The combination of any one of embodiments 22-33, wherein the signal sequence is a codon optimized signal sequence. 35. The combination of any one of embodiments 1-34, wherein one or more of said RNAs further comprise a poly-A tail. 36. The combination of any one of embodiments 1-35, wherein one or more of said RNAs further comprise a cap. 37. The combination of embodiment 36, wherein the cap is an m7GpppG cap, 3′-O-methyl- m7GpppG cap, or anti-reverse cap analog. 38. The combination of any one of embodiments 1-35, wherein one or more of said RNAs further comprise a cap-independent translational enhancer. 322 11979815v1 P-627574-PC 39. The combination of any one of embodiments 1-38, wherein one or more of said RNAs further comprise 5′ and/or 3′ untranslated regions. 40. The combination of any one of embodiments 1-39, wherein one or more of said RNAs are encapsulated in or formulated with a nanoparticle, lipid, polymer, cholesterol, and/or cell penetrating peptide. 41. The combination of embodiment 40, wherein said nanoparticle is a liposomal nanoparticle. 42. The combination of any one of embodiments 1-41, wherein said HSV infection comprises an HSV-1 infection. 43. The combination of any one of embodiments 1-41, wherein said HSV infection comprises an HSV-2 infection. 44. The combination of any one of embodiments 3-43, wherein said latent HSV infection comprises a genital HSV infection, an oral HSV infection, or a neurological HSV infection. 45. The combination of any one of embodiments 1-44, wherein one or more of said compositions is formulated for intramuscular administration. 46. The combination of any one of embodiments 1-44, wherein one or more of said compositions is formulated for subcutaneous administration. 47. The combination according to any one of embodiments 1-44, wherein one or more of said compositions is formulated for intradermal administration. 48. The combination of any one of embodiments 1-44, wherein one or more of said compositions is formulated for intranasal, intravaginal, or intrarectal administration. 49. The combination of any one of embodiments 1-44 wherein one or more of said compositions is formulated for topical administration. 50. The combination of any one of the embodiments 1-49, wherein said combination comprises: (a) a first composition comprising a nucleoside modified RNA encoding a first HSV glycoprotein, (b) a second composition comprising a nucleoside modified RNA encoding a second HSV glycoprotein, and (c) a third composition comprising a nucleoside modified RNA encoding a third HSV glycoprotein. 51. The combination of any one of the embodiments 1-50, wherein each of the RNAs encoding the HSV glycoproteins are contained in a single composition. 323 11979815v1 P-627574-PC 52. The combination of any one of embodiments 1-50, wherein the first composition, second composition, and third composition are the same composition. 53. The combination of any one of embodiments 1-52, further comprising additional administrations of said compositions to be provided to a subject subsequent to the first administration. 54. The combination of any one of embodiments 1-52, further comprising one or more doses of each of the first composition, second composition, or third composition. 55. The combination of any one of embodiments 1-54, wherein said combination further comprises: (a) a first composition comprising HSV gD or an immunogenic fragment thereof, (b) a second composition comprising HSV gC or an immunogenic fragment thereof, and (c) a third composition comprising HSV gE or an immunogenic fragment thereof. 56. The combination of any one of embodiments 4-55, wherein said latent HSV infection is in the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or any combination thereof. 57. The combination of any one of embodiments 4-51, wherein said latent HSV infection is detectable by HSV titers or HSV DNA in the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or any combination thereof. 58. Use of a combination for treating or inhibiting Herpes Simplex Virus (HSV) oral mucosal infection, encephalitis infection, or any combination thereof; or an HSV-1 infection, wherein the combination comprises: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions. 59. Use of a combination for the manufacture of a medicament for treating or inhibiting Herpes Simplex Virus (HSV) oral mucosal infection, an HSV encephalitis infection, or any combination thereof; or an HSV-1 infection, wherein the combination comprises: 324 11979815v1 P-627574-PC (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions. 60. The use of embodiments 58 or 59, wherein the infection is a latent infection, a primary infection, or a secondary infection. 61. The use of any one of embodiments 58-60, wherein the infection is in the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or any combination thereof. 62. Use of a combination for preventing establishment of a latent Herpes Simplex Virus (HSV) infection in a subject, wherein the combination comprises: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions. 63. Use of a combination for the manufacture of a medicament for preventing establishment of a latent Herpes Simplex Virus (HSV) infection in a subject, wherein the combination comprises: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, 325 11979815v1 P-627574-PC wherein the use comprises administering one or more doses of one or more of the compositions. 64. Use of a combination for treating or inhibiting a Herpes Simplex Virus (HSV) infection of the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or a combination thereof in a subject, wherein the combination comprises: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions. 65. Use of a combination for the manufacture of a medicament for treating or inhibiting a Herpes Simplex Virus (HSV) infection of the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or a combination thereof in a subject, wherein the combination comprises: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions. [0556] The following examples are presented in order to more fully illustrate the preferred embodiments of the disclosure. They should in no way be construed, however, as limiting the broad scope of the disclosure. EXPERIMENTAL DETAILS SECTION EXAMPLE 1: MATERIALS AND EXPERIMENTAL METHODS [0557] Modified RNA expressing exemplary HSV-2 glycoproteins C, D and E (gC2/gD2/gE2) ectodomains. Exemplary modified RNA (encoding an immunogenic gC2 fragment (SEQ ID NO: 326 11979815v1 P-627574-PC 10), encoding an immunogenic gD2 fragment (SEQ ID NO: 4), and encoding an immunogenic gE2 fragment (SEQ ID NO: 16)) was prepared based on the DNA coding sequences that encode HSV-2 glycoprotein C (gC2; amino acids 27-426 from HSV-2 strain 333; SEQ ID NO: 11), glycoprotein D (gD2; amino acids 26-331 from HSV-2 strain 333; SEQ ID NO: 5), and glycoprotein E (gE2; amino acids 24-405 from HSV-2 strain 2.12; SEQ ID NO: 17). [0558] The exemplary modified RNA was incorporated into lipid nanoparticles (LNP) to prepare the following immunogens: (a) Poly C RNA in LNP; (b) a modified RNA encoding an exemplary immunogenic gC2 fragment in LNP; (c) a modified RNA encoding an exemplary immunogenic gD2 fragment in LNP; (d) a modified RNA encoding an exemplary immunogenic gE2 fragment in LNP; (e) modified RNA encoding exemplary immunogenic gC2, gD2, and gE2 fragments in LNP. [0559] Immunization groups were as follows: a) Controls (Poly C group): 10μg Poly C RNA/LNP divided into 4 aliquots and administered at 4 separate sites. b) Exemplary gD2 immunogenic fragment alone (gD2 group): 10μg RNA encoding an exemplary immunogenic gD2 fragment /LNP divided into 4 aliquots and administered at 4 separate sites. c) Individual trivalent (Trivalent-I group): 10μg RNA encoding an exemplary immunogenic gC2 fragment /LNP, 10μg RNA encoding an exemplary immunogenic gD2 fragment /LNP, 10μg RNA encoding an exemplary immunogenic gE2 fragment/LNP each divided into 2 aliquots and each given at 2 sites. d) Combined trivalent (Trivalent-C group): 10μg RNA encoding an exemplary immunogenic gC2 fragment & 10μg RNA encoding an exemplary immunogenic gD2 fragment & 10μg RNA encoding an exemplary immunogenic gE2 fragment combined into LNP and divided into 4 aliquots and given at 4 sites. [0560] Experimental Procedures. Hair was removed from the back of 6-8 week old BALB/c mice using an electric razor and Nair. Mice were bled prior to the first and second immunization and prior to intravaginal challenge. Two immunizations were performed intradermally at 28-day intervals. Intradermal immunizations were performed on the denuded backs. Five mice that received the trivalent vaccine at individual sites (group c above) were sacrificed 14 days after the second immunization. Spleens were harvested for CD4+ and CD8+ T cell responses to exemplary gC2, gD2 and gE2 subunit immunogenic antigen fragments or to 15 amino acid peptides each with 327 11979815v1 P-627574-PC 11 overlapping amino acids. Twenty-eight days after the second immunization, mice were treated subcutaneously with 2mg Depo-Provera and 5 days later infected intravaginally with 5x103 PFU HSV-2 strain MS (~400 LD50). On days 2 and 4 post-challenge, vaginal swabs were obtained for virus cultures. On day 4 post-challenge, some mice in each vaccine group were sacrificed and dorsal root ganglia (DRG) excised for HSV-2 DNA qPCR. The remaining animals were evaluated for weight loss and hind limb weakness for 10 days while survival and genital disease were monitored for 28 days. EXAMPLE 2: CHARACTERIZATION OF TRANSLATIONAL PRODUCTS PRODUCED BY MODIFIED RNA ENCODING gC2, gD2, AND gE2 IMMUNOGENIC FRAGMENTS [0561] The ability of modified RNA to express proteins of the expected molecular weight when transfected into mammalian cells was verified.0.1μg of modified RNA encoding exemplary gC2, gD2, or gE2 immunogenic fragments was transfected into 293T cells using TransIT-mRNA (Mirus Bio LLC) for the transfection. Eighteen hours later, cells were harvested and extracts prepared for Western blots. The RNAs were designed to express immunogenic fragments of exemplary gC2, gD2 and gE2 ectodomain (labeled mRNA-ecto). As controls for the expected molecular weights, purified baculovirus proteins gC2, gD2, and gE2 expressing the same amino acids as the RNA constructs (labeled Bac-ecto) were used (Figures 1A-C). [0562] Conclusion: When transfected into mammalian cells, modified RNA encoding immunogenic fragments of HSV-2 gC2 (Figure 1A), gD2 (Figure 1B) and gE2 (Figure 1C) ectodomains produced proteins of the appropriate molecular weights that reacted with antibodies to the glycoproteins on Western blot. EXAMPLE 3: ELISA ANTIBODY RESPONSES IN SUBJECTS IMMUNIZED WITH gD2 IMMUNOGENIC FRAGMENT OR TRIVALENT MODIFIED RNA VACCINES [0563] ELISA endpoint titers were evaluated on sera taken 28 days after the first and second immunizations. Immunization groups were as follows: Poly C (10μg Poly C RNA/LNP divided into 4 aliquots and administered at 4 separate sites) (control); gD2 (10μg RNA encoding exemplary gD2 immunogenic fragment/LNP divided into 4 aliquots and administered at 4 separate sites); Trivalent-I (10μg RNA encoding exemplary gC2 immunogenic fragment /LNP, 10μg RNA encoding exemplary gD2 immunogenic fragment /LNP, 10μg RNA encoding exemplary gE2 328 11979815v1 P-627574-PC immunogenic fragment /LNP each divided into 2 aliquots and each given at 2 sites); and Trivalent-C (10μg RNA encoding exemplary gC2 immunogenic fragment & 10μg RNA encoding exemplary gD2 immunogenic fragment & 10μg RNA encoding exemplary gE2 immunogenic fragment combined into LNP and divided into 4 aliquots and given at 4 sites). [0564] Four animals were evaluated in each group. High ELISA titers to each immunogen were obtained after the first immunization (marked as roman numeral I; Figures 2A-C), and the titers were boosted even higher after the second immunization (marked as roman numeral II; Figures 2A-C). Immunization with gD2 vaccine selectively induced extremely high titers of ELISA antibodies to gD2 (Figure 2B), while immunization with the trivalent modified RNA vaccines induced extremely high titers of ELISA antibodies to gC2 (Figure 2A) and gD2 (Figure 2B) and high titers to gE2 (Figure 2C). In all non-control groups, the second immunization significantly boosted the ELISA titers compared to the first. The differences between the titers in the second vs the first immunizations were significant, p<0.05 (t-tests, comparing the antibody titers after the first and second immunization). [0565] Conclusion: The RNA vaccine encoding gD2 and the trivalent RNA vaccines induced extremely high titers of ELISA antibodies after the first immunization that were significantly boosted after the second immunization. EXAMPLE 4: BALANCED TH1 AND TH2 IgG ISOTYPES PRODUCED BY MODIFIED RNA IMMUNIZATION [0566] The ability of RNA immunizations to stimulate predominantly a TH1 or TH2 immune response was tested by determining whether IgG1 (TH2) or IgG2a (TH1) antibodies are produced. ELISA was performed on plates coated with all three antigens, gC2, gD2 and gE2. Serum obtained after the first or second immunization was added to the antigen-coated plates, and IgG1 or IgG2a was detected using HRP anti-mouse IgG1 or IgG2a. IgG1 (Figure 3A) and IgG2a (Figure 3B) titers were significantly elevated after immunization with the gD2 vaccine and the trivalent modified RNA vaccines. Further, the IgG1 (Figure 3A) or IgG2a (Figure 3B) titers were significantly higher after the second modified RNA immunization compared to the first, p<0.05 (t tests). [0567] Conclusion: The results demonstrate high titers of antibodies are produced to both IgG1 and IgG2a isotypes, indicating a balanced TH1 and TH2 response to immunization with modified RNA encoding exemplary gC2, gD2 and gE2 immunogenic fragments. 329 11979815v1 P-627574-PC EXAMPLE 5: HIGH NEUTRALIZING ANTIBODY TITERS AFTER MODIFIED RNA IMMUNIZATION [0568] Serum was obtained 28 days after the second immunization, and neutralizing antibody titers were determined using serial 2-fold dilutions of serum, starting at a 1:25 dilution and 10% human serum as a source of complement. The human serum was obtained from an individual seronegative for HSV-1 and HSV-2. The modified RNA groups were each significantly different from the poly C controls (p<0.001; Figure 4). While each of the RNA groups was not significantly different from one another, the trivalent vaccine given as a combined immunogen (Trivalent-C) performed the best of the three RNA groups (Figure 4). [0569] Conclusion: Each of the modified RNA groups produced extremely high titers of neutralizing antibodies in the presence of 10% human complement. EXAMPLE 6: CD4+ AND CD8+ T CELL RESPONSES IN SPLENOCYTES AFTER MODIFIED RNA IMMUNIZATION [0570] Five animals from the trivalent modified RNA group that were immunized with each exemplary glycoprotein immunogenic fragment RNA at a separate site (Trivalent-I group) were euthanized 14 days after the second immunization. Splenocytes were prepared for T cell assays. Splenocytes were stimulated with glycoprotein subunit antigens prepared in baculovirus or 15 amino acid peptides containing 11 overlapping amino acids. The CD4+ and CD8+ T cell responses are shown in Figures 5A-5B and Figures 6A-6B, respectively. [0571] CD4+ T cells: The modified RNA-expressed exemplary gC2, gD2, and gE2 subunit immunogenic fragments each stimulated polyfunctional CD4+ T cell responses (Figures 5A-5B). Splenocytes harvested from immunized subjects and then stimulated with subunit glycoprotein immunogenic fragments increased polyfunctional CD4+ T cell responses (Figure 5A). Splenocytes harvested from immunized subjects and then stimulated with 15 amino acid overlapping peptides increased polyfunctional CD4+ T cell responses and IFNγ responses (Figure 5B). [0572] CD8+ T cells: Only exemplary gE immunogenic peptide pool 2 stimulated a significant IFNγ CD8+ T cell response (Figures 6A-6B). 330 11979815v1 P-627574-PC EXAMPLE 7: SURVIVAL, WEIGHT LOSS AND NEUROLOGICAL SIGNS AFTER MODIFIED RNA IMMUNIZATION AND INTRAVAGINAL CHALLENGE [0573] Thirty-three days after the second immunization, animals were inoculated intravaginally with 5x103 PFU of HSV-2 strain MS (~400 LD50). Animals were observed daily for survival, neurological signs consisting of hind limb weakness or paralysis and hunched gait, and for weight loss or gain. All animals in the poly C control group died, while all animals in the gD2 vaccine alone, trivalent given individually (labeled Trivalent-I) or trivalent given combined (labeled Trivalent-C) survived (Figure 7A; p=0.002 by Log-rank (Mantel-Cox) comparing the three exemplary RNA/LNP groups with poly C controls). Figure 7B demonstrates that administration of the modified RNA vaccine twice at 28 day intervals and challenged intravaginally with HSV-2 does not result in neurological signs or weight loss. Control subjects that were not administered the vaccine and were challenged intravaginally with HSV-2 showed weight loss and neurological signs. [0574] Each of the RNA/LNP groups significantly outperformed the control group. All mice immunized with the modified RNA survived and showed no evidence of weight loss, neurological disease or genital lesions after intravaginal challenge with ~400 LD50 of HSV-2. EXAMPLE 8: HSV-2 VAGINAL TITERS AFTER MODIFIED RNA IMMUNIZATION AND INTRAVAGINAL CHALLENGE [0575] Vaginal swabs were obtained from 10 animals per group on days 2 and 4 post challenge and cultured for replication competent HSV-2 virus. Results are shown in Figures 8A-8B. 9/10 animals in the poly C group had positive cultures on days 2 (Figure 8A) and 4 (Figure 8B) compared with 3/10 in the gD2 vaccine group and 0/10 in the trivalent-I or trivalent-C groups (P values by Fisher Exact test were not significant comparing trivalent groups to gD2 vaccine group; p<0.001 comparing trivalent-I or trivalent-C with poly C; p=0.02 comparing gD2 vaccine with poly C). [0576] Each of the RNA/LNP groups significantly outperformed the Poly C control group. Remarkably, day 2 and day 4 vaginal titers after challenge were negative in mice immunized with the trivalent RNA whether given at separate sites or as a combined immunization. No significant differences were detected comparing either the trivalent group with the gD2 vaccine group, although both trivalent groups outperformed the gD2 group, as 3 of 10 mice in the gD2 group had virus isolated from vaginal swabs. 331 11979815v1 P-627574-PC EXAMPLE 9: GENITAL DISEASE AFTER MODIFIED RNA IMMUNIZATION AND INTRAVAGINAL CHALLENGE [0577] Animals were monitored daily for genital disease for 28 days post challenge. A score of 0 was assigned for no disease, and 1 point each was assigned for hair loss around the anal or genital orifices, genital erythema, genital exudate, and necrosis of genital tissues (Figure 9). [0578] No animal in the gD2 or trivalent RNA/LNP groups developed genital disease, which was significantly different than the poly C controls (p<0.001, one-way ANOVA by Kruskal-Wallis test followed by Dunn’s multiple comparisons for significance). EXAMPLE 10: HSV-2 DNA IN DORSAL ROOT GANGLIA AFTER MODIFIED RNA IMMUNIZATION AND INTRAVAGINAL CHALLENGE [0579] Five animals per group were euthanized at 4 days post challenge, except for the trivalent- combined group in which four animals were euthanized. Dorsal root ganglia (DRG) were harvested for HSV-2 DNA quantitation by qPCR to detect the Us9 gene. All five animals in the poly C group had HSV-2 DNA detected in the DRG, while 2/5 animals in the gD2 group, 1/5 in the trivalent RNA administered at individual sites group (Trivalent-I), and 1/4 in the trivalent modified RNA given at the same site group (Trivalent-C) were positive for HSV-2 DNA (Figure 10; Mann-Whitney test: gD2 group compared with poly C, p=0.03; trivalent at different sites compared with poly C, p<0.01; Trivalent-I compared with poly C, p=0.14). The difference between the modified RNA immunized groups was not significant. [0580] Summary: Dorsal root ganglia were negative for HSV-2 DNA on day 4 after infection in 75% to 80% of animals immunized with modified RNA encoding gD2 immunogenic fragment alone or the trivalent vaccine. The trivalent RNA group administered individually at different sites and the gD2 groups significantly outperformed the poly C RNA control group, while the trivalent RNA group administered together did not differ significantly from the poly C group, likely because of the smaller sample size in the trivalent-combined group. [0581] Modified RNA vaccines expressing gD2 immunogenic fragment alone or gC2, gD2 and gE2 (trivalent) immunogenic fragments provided outstanding protection against HSV-2 genital challenge. The expression of the three exemplary immunogenic fragments slightly outperformed exemplary gD2 immunogenic fragment based on day 2 and day 4 titers after challenge and the lower number of animals with HSV-2 DNA detected in DRG on day 4. 332 11979815v1 P-627574-PC EXAMPLE 11: T FOLLICULAR HELPER (Tfh) CELL AND GERMINAL CENTER B CELL RESPONSES IN IMMUNIZED MICE [0582] BALB/c female mice were left un-immunized as naïve control animals or immunized intradermally twice at 28 day intervals with poly C RNA/LNP or trivalent modified RNA-LNP. The poly C RNA controls received 10μg Poly C RNA-LNP divided into 4 aliquots and administered at 4 separate sites. The trivalent modified RNA group received 10μg RNA encoding exemplary gC2 immunogenic fragment/LNP, 10μg modified RNA encoding exemplary gD2 immunogenic fragment/LNP, and 10μg modified RNA encoding exemplary gE2 immunogenic fragment/LNP each divided into 2 aliquots and each given at 2 sites. Two weeks after the second immunization, spleens were harvested from 5 animals per group and flow cytometry performed to detect T follicular helper (Tfh) cells (Figure 11A; *p<0.05) and germinal center B cell responses (Figure 11B; *p<0.05). [0583] Conclusion: The trivalent modified RNA/LNP vaccine induced a potent Tfh and germinal center B cell response and significantly outperformed the poly C control immunization (p<0.05) and the naïve group (p<0.05) for both Tfh and germinal center B cell responses. These immune responses suggest that the trivalent modified RNA/LNP vaccine will likely induce a durable antibody response. EXAMPLE 12: VAGINAL IgG RESPONSES TO MODIFIED RNA IMMUNIZATION IN MICE [0584] BALB/c mice were immunized intradermally twice at 28 day intervals with 10μg of poly C RNA-LNP, 10μg modified RNA encoding exemplary gD2 immunogenic fragment/LNP or 10μg trivalent modified RNA encoding each of exemplary gC2, gD2, and gE2 immunogenic fragments/LNP. The trivalent modified RNA was combined and administered as 10μg modified RNA encoding exemplary gC2 immunogenic fragment, 10μg modified RNA encoding exemplary gD2 immunogenic fragment, and 10μg modified RNA encoding exemplary gE2 immunogenic fragment combined into LNP and divided into 4 aliquots and given at 4 sites. One month after the second immunization, 60μl of media were introduced in the vaginal cavity and retrieved. IgG titers to gC2 (Figure 12A), gD2 (Figure 12B), and gE2 (Figure 12C) were determined at a 1:50 dilution of the vaginal wash fluids by ELISA (Figures 12A-12C, n=10 mice in the poly C group, n=10 in the gD2 modified mRNA group and n=25 in the trivalent modified mRNA group; ***p<0.001; **p<0.01). 333 11979815v1 P-627574-PC [0585] Conclusion: The trivalent modified RNA produced a robust vaginal IgG response to gC2 (Figure 12A) and gD2 (Figure 12B) and a more moderate response to gE2 (Figure 12C). The gD2 ELISA titers were higher in mice immunized with the trivalent RNA vaccine compared to mice immunized with the gD2 RNA vaccine (Figure 12B). EXAMPLE 13: ANTIBODIES TO EXEMPLARY gC2 IMMUNOGENIC FRAGMENT PRODUCED BY TRIVALENT RNA IMMUNIZATION OF MICE BLOCK IMMUNE EVASION DOMAINS ON gC2 [0586] BALB/c mice were left unimmunized as a source of non-immune IgG, or immunized intradermally with poly C RNA/LNP or trivalent modified RNA/LNP. The poly C RNA controls received 10μg poly C RNA/LNP divided into 4 aliquots and administered at 4 separate sites. The gD2 group received 10μg modified RNA encoding exemplary gD2 immunogenic fragment/LNP administered as described for the poly C RNA/LNP. The trivalent modified RNA group received 10μg modified RNA encoding exemplary gC2 immunogenic fragment/LNP, 10μg modified RNA encoding exemplary gD2 immunogenic fragment/LNP, and 10μg modified RNA encoding exemplary gE2 immunogenic fragment/LNP combined into one LNP and divided into 4 aliquots and given at 4 sites. There were 10 mice in each group. Sera from the 10 mice were pooled and IgG was purified. The IgG was evaluated at 12μg/200μl for its ability to block complement component C3b binding to gC2. This blocking assay is used to assess whether antibodies produced by immunization block the immune evasion properties of gC2. Non-immune murine IgG, IgG from the poly C RNA group, and IgG from the gD2 group each failed to block gC2 binding to C3b. In contrast, IgG from trivalent modified RNA-immunized animals totally blocked the interaction between gC2 and C3b (Figure 13, ****p<0.0001). [0587] Conclusions: The trivalent modified RNA vaccine produces antibodies that block immune evasion domains on gC2 as determined by blocking the interaction between gC2 and C3b. EXAMPLE 14: INTRAVAGINAL INFECTION OF MICE AT A HIGHER INOCULUM TITER OF HSV-2 AFTER MODIFIED RNA VACCINATION [0588] BALB/c mice (n=5) were immunized with the trivalent modified RNA using 10μg modified RNA encoding exemplary gC2 immunogenic fragment/LNP, 10μg modified RNA encoding exemplary gD2 immunogenic fragment/LNP, 10μg modified RNA encoding exemplary gE2 immunogenic fragment/LNP each divided into 2 aliquots and each given individually at 2 334 11979815v1 P-627574-PC sites. One month after the second immunization, mice were treated with medroxyprogesterone and five days later infected intravaginally with 5x104 PFU HSV-2 strain MS (2,000 LD50). Animals were followed for 28 days and evaluated for death, genital disease, vaginal viral titers 2 and 4 days after infection and dorsal root ganglia (DRG) HSV-2 DNA copy number 28 days after infection. No mouse immunized with the trivalent modified RNA/LNP vaccine died, had genital disease, had any virus detected on day 2 or 4 post-infection or had HSV-2 DNA detected in DRG (Table 8). Table 8. Trivalent modified RNA/LNP immunized mice challenged with HSV-2 strain MS (2,000 LD50) Disease parameters Mice % Protection Death 0/5 100 [0589] Co
Figure imgf000337_0001
c us o s: ce were n ec e w - a a ose a was - o gher than used in earlier experiments described herein (Figures 7A-10). Protection of the mice remained outstanding even at this higher titer challenge. All five mice achieved sterilizing immunity as determined by no deaths, no genital disease, negative vaginal virus titers on days 2 and 4 post- infection and no HSV-2 DNA in the lumbosacral DRG on day 28 (Table 8). EXAMPLE 15: EVALUATION OF THE INTRAMUSCULAR ROUTE OF MODIFIED RNA IMMUNIZATION IN MICE [0590] BALB/c mice were immunized intramuscularly with poly C RNA/LNP as a control (15/group) or with trivalent modified RNA containing 10μg of modified RNA encoding each of exemplary gC2, gD2, and gE2 immunogenic fragments/LNP (20/group). All poly C control animals died by day 12, while all animals in the trivalent modified RNA group survived (Figure 14A). No weight loss occurred in the trivalent modified RNA group, while the poly C control animals lost >15% of body weight (Figure 14B). The poly C group developed extensive genital disease, while the trivalent modified RNA animals had no genital disease (Figure 14C). DRG were harvested from nine poly C animals at the time of euthanasia between days 7 and 12 post- infection or at the end of the experiment on day 28 in the trivalent modified RNA group. All 335 11979815v1 P-627574-PC animals in the poly C group had HSV-2 DNA detected in DRG, while none were positive for HSV- 2 DNA in trivalent modified RNA group (Figure 14D). Day 2 (Figure 14E) and Day 4 (Figure 14F) vaginal viral cultures were positive in all 15 animals in the poly C group, while cultures were negative in all 20 animals in the trivalent modified RNA group. Differences between poly C and trivalent groups are significant, p<0.001 for each figure (Figures 14A-14F). [0591] Conclusions: Trivalent modified RNA/LNP provides outstanding protection in mice when administered intramuscularly. Comparable findings to the above were observed when mice were immunized intradermally. Overall, 64 mice have been evaluated, which were immunized with trivalent modified RNA at 10μg of each immunogenic fragment given either intradermally (Figures 7A-10) or intramuscularly (Figures 14A-14F). Sterilizing immunity was achieved in 63/64 (98%) mice based on no death, no genital disease, no weight loss, negative day 2 and 4 vaginal titers and negative HSV-2 DNA in DRG. EXAMPLE 16: SUMMARY COMPARISON OF IMMUNIZATION WITH TRIVALENT RNA/LNP AND TRIVALENT SUBUNIT ANTIGEN CPG/ALUM IN BALB/c MICE [0592] The results presented in Table 9 hereinbelow represent a summary of all the results in BALB/c mice that were immunized either intradermally or intramuscularly with trivalent modified RNA containing 10μg of modified RNA encoding gC2, gD2 and gE2 immunogenic fragments/LNP (total 64 mice studied). A comparison is shown with the results obtained in BALB/c mice that were immunized with 5μg each of bac-gC2(27-426t) containing gC2 amino acids 27-426 from HSV-2 strain 333, bac-gD2(306t) containing gD2 amino acids 26-331 from HSV-1 strain 333, and bac-gE2(24-405t) containing gE2 amino acids 24-405 from HSV-2 strain 2.12. The exemplary gC2, gD2, gE2 fragment antigens were mixed with 150μg CpG and 25μg alum/per μg protein as adjuvants and administered intramuscularly. Mice were immunized twice at 28-day intervals with trivalent modified RNA/LNP and three times at 14-day intervals with exemplary gC2, gD2, gE2 fragment antigens, as done in prior experiments. The modified RNA and gC2, gD2, gE2 fragment antigen experiments were performed at the same time. The results summarized in Table 9 demonstrate significant superiority of the trivalent modified RNA/LNP vaccine over the trivalent gC2, gD2, gE2 fragment antigen vaccine in many immune response parameters, and most importantly in vaccine efficacy. The trivalent modified RNA/LNP vaccine achieved sterilizing immunity in 63/64 (98%) of mice compared to 15/20 (75%) in the subunit antigen group. 336 11979815v1 P-627574-PC Table 9. Comparisons of immunization with trivalent modified RNA/LNP or trivalent exemplary gC2, gD2, gE2 fragment antigen CpG/alum in BALB/c mice. Comparison Trivalent modified Trivalent P value RNA exemplary gC2, A A
Figure imgf000339_0001
337 11979815v1 P-627574-PC *NS, not significant; #Sterilizing immunity defined as no death, no genital disease, no weight loss, and no evidence of subclinical infection as measured by day 2 and day 4 vaginal cultures post- infection and HSV-2 DNA in dorsal root ganglia on day 4 or day 28 post-infection. EXAMPLE 17: EVALUATION OF THE TRIVALENT MODIFIED RNA/LNP VACCINE IN GUINEA PIGS [0593] Hartley Strain female guinea pigs were left unimmunized and uninfected (naive group, n=10), immunized three times intradermally at monthly intervals with 20μg poly C RNA-LNP (n=10) or with 20μg of modified RNA encoding each of exemplary gC2, gD2, gE2 immunogenic fragments/LNP (n=10). One month after the final immunization, animals in the poly C and trivalent modified RNA groups were infected intravaginally with 5x105 PFU of HSV-2 strain MS (50 LD50). Animals were observed for death, genital lesions during the acute phase of infection (days 1-14) and genital lesions during the recurrent phase of infection (days 15-60). In the poly C control group, 7/10 animals died or were humanely euthanized between days 7 and 20 post- infection, while no animal in the trivalent group and no naïve (uninfected) animal died (Figure 15A). The poly C group had genital lesions on a mean of 6.4 days during the acute phase of infection with 9/10 animals developing acute genital disease, while no animal in the trivalent group or naïve (uninfected) group developed acute genital disease (Figure 15B). The poly C animals had genital lesions on a mean of 3.7 days during the recurrent phase of infection from days 15-60, with 2/3 animals developing recurrent genital lesions (Figure 15C). In contrast, the trivalent immunized guinea pigs and the naïve (uninfected) animals had no recurrent genital lesions (Figure 15C). [0594] Conclusions: Trivalent modified RNA/LNP provided outstanding protection against acute and recurrent genital lesions in guinea pigs. EXAMPLE 18: EVALUATION OF THE HSV-1 AND HSV-2 TRIVALENT NUCLEOSIDE-MODIFIED RNA/LNP VACCINES AGAINST GENITAL AND NON- GENITAL HSV-1 INFECTION IN MICE [0595] The present Example demonstrates that an RNA vaccine encoding exemplary HSV gC, gD, and gE immunogenic fragments is effective against genital HSV-1 and HSV-2 infection, as well as HSV-1 oral infection. Further, the present Example demonstrates the unexpected finding that an RNA vaccine encoding HSV-2 gC, gD, and gE (“tri-HSV-2”) showed similar efficacy against HSV-1 as an RNA vaccine encoding HSV-1 gC, gD, and gE (“tri-HSV-1”). 338 11979815v1 P-627574-PC Materials and Methods [0596] Tri-HSV-1 and tri-HSV-2 modified RNA encoding exemplary immunogens. The tri- HSV-2 immunogens were prepared to generate RNA that encodes exemplary immunogenic fragments of the ectodomains of HSV-2 gC (27-426; SEQ ID NO: 11), HSV-2 gD (26-333; SEQ ID NO: 120), and HSV-2 gE (24-405; SEQ ID NO: 17). The tri-HSV-1 exemplary immunogens were prepared to generate RNA that encodes exemplary immunogenic fragments of the ectodomains of the HSV-1 gC (27-457; SEQ ID NO: 8), HSV-1 gD (26-333; SEQ ID NO: 134), and HSV-1 gE (24-309; SEQ ID NO: 140) proteins where the first amino acid listed represents the amino acid immediately after the signal sequence. Each subunit protein extends from the first amino acid after the signal sequence to just prior to the transmembrane domain. [0597] To generate nucleoside-modified RNA, m1Ψ-5′-triphosphate (TriLink) was used instead of UTP. RNA was capped using the m7G capping kit with 2′-O-methyltransferase (ScriptCap, CellScript). The RNA contains a 101-nucleotide poly(A) tail. Nucleoside-modified RNAs were purified by Fast Protein Liquid Chromatography (FPLC) (Akta Purifier GE Healthcare) and stored at -20°C. FPLC nucleoside-modified RNAs and polyC RNA (Sigma) were encapsulated in LNPs using a self-assembly process in which an aqueous solution of RNA at acidic pH 4.0 was rapidly mixed with a solution of lipids dissolved in ethanol. LNPs used in this study were similar in composition to those described previously, which contain an ionizable cationic lipid/phosphatidylcholine/cholesterol/PEG-lipid (Acuitas) (50:10:38.5:1.5 mol/mol) and were encapsulated at an RNA to total lipid ratio of ~0.05 (wt/wt). The LNPs had a diameter of ~80 nm as measured by dynamic light scattering using a Zetasizer Nano ZS (Malvern Instruments Ltd, Malvern, UK) instrument, and were stored at -80°C at a concentration of RNA of ~1 μg/μl. [0598] The three modified RNAs encoding exemplary HSV-1 immunogenic fragments were combined in equal concentrations based on mass prior to encapsulation in lipid nanoparticles by Acuitas. The vaccine formulations were stored at -80 ℃ and not reused once thawed. [0599] Immunization of mice. Female BALB/c mice aged 6-8 weeks old were immunized twice 30 days apart in the hind right gastrocnemius muscle. Mice were immunized with either 1 µg or 10 µg (total) of tri-HSV-1 or tri-HSV-2 modified RNA vaccines. Control mice were injected with an equal volume of sterile saline. Serum was collected 25 days after the final immunization and stored at -80 ℃. [0600] Serum IgG ELISA endpoint titers. Purified baculovirus-derived exemplary gC1, gD1, and gE1 immunogenic fragments were produced as previously described. Purified protein was added 339 11979815v1 P-627574-PC to High Binding Costar microtiter plates. Serial 2-fold dilutions of serum starting at 1:250 were added to protein coated wells. Anti-mouse IgG was detected by adding 2,2’-azino-bis-3- ethylbenzothiazoline-6-sulfonic acid (ABTS). Endpoint titers were calculated as the serum concentration that had an absorbance value of ^ 0.1 and was ^ two-fold higher than serum from PBS immunized mice. [0601] HSV-1 and HSV-2 neutralizing antibody titers. Neutralizing titers were measured by incubating 2-fold serial dilutions of mouse sera with 100 PFU of HSV-1 strain NS at 37℃ for 1 hour. Incubations were done in the presence of 5% human serum as a source of complement obtained from an HSV-1/HSV-2 seronegative donor. Remaining infectious virus was determined by plaque assay on Vero cells. The neutralizing titer was defined as the highest serum dilution that reduced plaques by ≥ 50%. [0602] Isolation and stimulation of splenocytes from BALB/c mice. Mice were immunized twice with PBS or 10 µg of exemplary tri-HSV-1 or tri-HSV-2 vaccine. Immunized mice were sacrificed 10 days after the second immunization. The spleens were harvested, and mechanically dissociated into single-cell suspensions. Cell counts were obtained and 2×106 cells were added to individual wells for each stimulation condition for each mouse. Splenocytes were stimulated in RPMI media supplemented with 55 µM 2-mercaptoethanol and 10 units recombinant mouse IL-2/µL stimulation media. Intracellular cytokines were immobilized with 5 µg/mL Brefeldin A and Monensin. Cells were stimulated with Dimethyl Sulfoxide (DMSO), 0.04 µM phorbol-12- myristate 13-acetate (PMA) with 0.67 µM ionomycin, or 1 µg/mL of HSV-1 protein overlapping peptide pools. Cells were incubated for 12 hours at 37 ℃ in 5% CO2. [0603] Overlapping peptide pools. Overlapping peptide pools were synthesized by GenScript with a purity of ≥70%. Peptides were 15 amino acids long with an 11 amino acid overlap. Identical amino acid sequences used in the modified RNA encoding exemplary immunogenic fragments were used for synthesizing overlapping peptide sequences. All peptide pools were resuspended in DMSO and RPMI to a concentration of 2 µg/µL and stored at -80 ℃. [0604] Flow cytometry reagents. All antibodies were purchased from BioLegend. Antibodies used for labeling cells were fluorescein isothiocyanate (FITC) conjugated anti-CD3 (clone 17A2), allophycocyanin (APC)-Fire 750 conjugated to CD4 (Clone GK1.5), phycoerythrin (PE) conjugated anti-IL-2 (cloneJES6-5H4), PE-Dazzle 594 conjugated anti-CD107a (clone 1D4B), Peridinin-Chlorophyll-Protein-Cyanin (PerCP-Cy5.5) conjugated anti-CD8α (clone 53-6.7), Alexa-Flour (AF)-700 conjugated anti-IFNγ (clone XMG1.2), Brilliant Violet 421 conjugated 340 11979815v1 P-627574-PC anti-IL-4 (Clone 11b11) and PE-Cy7 conjugated anti-TNFα (clone MP6-XT22). Dead cells were discriminated by Fixable Live-Dead Aqua dye (Invitrogen). Brefeldin A, PMA/ionomycin, Cyto- fast-fix-perm buffer, and intracellular staining permeabilization wash buffer were purchased from BioLegend. After labeling of intracellular cytokines, cells were fixed with 1% formaldehyde in PBS and stored at 4℃ until the next day. Cells were analyzed on a BD LSR 2. Gates were set to discriminate singlet cells and then lymphocytes based on forward scatter and side scatter properties. 150,000 events in the lymphocyte gate were collected. After data collection, flow samples were analyzed using FlowJo version 10.8 (Tree Star software). Cells were gated on CD4+ or CD8+ T-cells producing single cytokines. Boolean combination gates were used to compare IL- 2, TNFα, and IFNγ production. [0605] HSV-1 lip infection. Mice were challenged 30 days after the 2nd immunization. Mice were anesthetized with a solution of 100 mg/kg Ketamine and 12.5 mg/kg Xylazine, placed on their back, and a 25G needle was used to make 10 vertical scratches on the lower lip. After scratching the lip, 2×106 PFU of HSV-1 strain NS was topically applied. Mice were monitored for 13 days for weight loss and presence of lesions on the lower lip. [0606] Lip and Trigeminal (TG) tissue isolation. The lip and TG were removed and placed in tubes containing 500 µL DMEM supplemented with 5% FBS. The tissues were stored at -80 ℃ until further processing. Lip and TG tissues were subjected to 3 freeze-thaw cycles. The lip was homogenized in a glass grinder and resuspended in 1 mL of media. The TG was homogenized using the frosted ends of glass slides and resuspended in 275 µL of DMEM. Tissue homogenates were used to make 10-fold serial dilutions that were then evaluated by plaque assay on Vero cells. [0607] HSV-1 vaginal infection. Mice were challenged with 2×106 PFU of HSV-1 strain NS intravaginally. Mice were scored daily for 14 days on a basis of one point each for redness/erythema, hair loss, distended belly indicating loss of fecal motility, and urinary staining for a maximum daily score of four. The vaginal canals of mice were swabbed on days 2 and 4 for virus titers. Vaginal swabs were placed in 1 mL of DMEM supplemented with L-glutamine, HEPES, antibiotics, and 5% FBS. Serial 10-fold dilutions were evaluated by plaque assay on Vero cells. [0608] HSV-1 DNA isolation. The DRG were harvested at the time of sacrifice of the mice either at the end of the experiment on day 28 post-challenge or prior to day 28 in mice in which humane euthanasia was required. Purified DNA was isolated and amplified for qPCR as previously described and the HSV-1 DNA copy number was expressed as log10 DNA copies per 105 adipsin 341 11979815v1 P-627574-PC genes. Samples with less than one copy by 40 cycles in duplicate wells were considered negative. If only one of the duplicate wells was positive, the sample was tested in triplicate and considered positive if two or more of the triplicates were positive. [0609] Statistics. The log-rank test was used to calculate P values for survival curves. The Mann- Whitney test was used to calculate P values in experiments evaluating vaginal titers, serum antibodies by ELISA, HSV-1 neutralizing titers, viral titers present in lip and TG tissue, and HSV- 1 DNA copy number in the lip, TG, and DRG. The flow cytometry data was analyzed using 2-way ANOVA with correction for multiple comparisons. All significance tests were two-tailed with a P value < 0.05 considered significant. Analysis was done using GraphPad Prism version 9.5 (GraphPad software Inc). Results [0610] The efficacy of exemplary tri-HSV-1 vaccine and exemplary tri-HSV-2 vaccine against HSV-1 challenge to lip or vagina was compared. To better resolve differences between the vaccines, mice were immunized with either a 1 µg (low) or 10 µg (high) dose of total modified RNA containing equal concentrations of each modified RNA immunogen. The tri-HSV-1 immunized mice had significantly higher serum IgG ELISA antibodies to gC1 and gE1 than tri- HSV-2 immunized animals (Figures 16A-16B). The gC1 IgG levels were 4.6-fold higher in the tri-HSV-1 group than in the tri-HSV-2 group at 1 µg and 6.9-fold higher at 10 µg doses. A dose response with the tri-HSV-1 vaccine was detected in that the gC1 IgG levels were 2.3-fold higher in animals immunized with 10 µg than 1 µg, while no differences were detected at the two doses in the tri-HSV-2 immunized mice (Figure 16A). The gE1 IgG levels were 35- and 72-fold higher in tri-HSV-1 than tri-HSV-2 immunized mice at the low and high doses, respectively. A 2.5-fold dose-response increase in gE1 IgG titers in the high dose tri-HSV-1 immunized mice (Figure 16B) was detected. The gD1 IgG levels were not significantly different comparing tri-HSV-1 and tri- HSV-2 immunized mice at the low or high dose (Figure 17C). The IgG ELISA results suggest that many epitopes on gD1 and gD2 are type common, while epitopes on gC1 and gC2, or gE1 and gE2 are type specific. [0611] Sera was evaluated for HSV-1 neutralizing antibodies in the presence of 5% human serum as a source of complement. Neutralizing titers were higher in the tri-HSV-1 vaccine immunized mice at both immunization doses, but differences did not reach statistical significance (Figure 16D). The neutralizing antibody results are consistent with the gD ELISA results showing comparable IgG binding titers to gD1 produced by the tri-HSV-1 and tri-HSV-2 vaccines. The 342 11979815v1 P-627574-PC neutralizing titers are also consistent with the prior observation that gD antibodies are more potent at neutralizing virus than gC or gE antibodies. Tri-HSV-2 immunization induces a more robust CD8+ T-cell responses to gE1 overlapping peptides than tri-HSV-1 immunization. [0612] The T-cell response in mice immunized twice with 10 µg tri-HSV-1 or tri-HSV-2 vaccines was evaluated. Splenocytes were stimulated with overlapping peptide pools of gC1, gD1, or gE1. Responding cells were measured by intracellular cytokine flow cytometry (Figure 17). CD8+ T cell responses to gC1 or gD1 in tri-HSV-1 or tri-HSV-2 immunized mice were not detected (Figure 18A). However, robust CD8+ T cell responses to gE1 overlapping peptides was detected, particularly in mice immunized with the tri-HSV-2 vaccine (Figure 18A). The number of IFNγ+TNFα+ (double positive) CD8+ T-cells was 3.4-fold higher in the tri-HSV-2 immunized mice than the tri-HSV-1 immunized vaccine group (Figure 18B, 3rd group from right). Additionally, the number of triple positive CD8+ T-cells (IFNγ+TNFα+IL-2+) was 8.2-fold higher in the tri- HSV-2 vaccine immunized group compared to the tri-HSV-1 vaccine immunized group (Figure 18B, last group on right). CD8+ T-cell activity was analyzed by CD107a labeling, a marker of degranulation. The tri-HSV-1 vaccine immunized group had more activated T-cells compared to the PBS group, but the tri-HSV-2 vaccine immunized animals had 7-fold greater CD107a labeling compared to the tri-HSV-1 vaccine immunized animals (Figure 18C). CD4+ T-cells stimulated with gD1 overlapping peptides were analyzed. CD4+ T-cells in the tri-HSV-2 vaccine immunized group had a 2-fold increase in polyfunctional cells (IFNγ+TNFα+IL-2+) compared to tri-HSV-1 vaccine immunized mice (Figure 18D). Responding cells were evaluated to determine whether the cells have a Th1 or Th2 phenotype by calculating the ratio of IFNγ+ (Th1) to IL-4+ (Th2) responses. A high ratio of IFNγ to IL-4 was observed, indicating a Th1 phenotype (Table 10). [0613] Table 10. Ratio of CD4+ T-cells producing IFNγ to IL-4 following stimulation with gD1 overlapping peptides. PBS 1.25
Figure imgf000345_0001
[0614] CD4+ T cell responses to gC1 or gE1 overlapping peptides in exemplary tri-HSV-1 or exemplary tri-HSV-2 vaccine immunized mice were not detected. These results indicate that immunization with exemplary tri-HSV-2 RNA vaccine induces CD4+ and CD8+ effector T-cell 343 11979815v1 P-627574-PC responses in mice to peptides from HSV-1, and that some CD4+ and CD8+ T cell responses to the exemplary tri-HSV-2 vaccine are even more potent than to the exemplary tri-HSV-1 vaccine. [0615] A challenge model that targets a non-genital site for infection was selected to compare efficacy of tri-HSV-1 and tri-HSV-2 modified RNA vaccine immunizations. The most common site of HSV-1 infection is in the oral mucosa. The lip scarification model allows virus to be added to the lower lip adjacent to the oral mucosa. Scratch inoculation of the virus on the lower lip causes productive infection in the lip and trigeminal ganglia (TG) infection via HSV-1 transport along the mandibular branch of the trigeminal nerve. Mice were immunized and later infected by scratch inoculation of 2×106 PFU of HSV-1 strain NS to the lower lip. PBS-immunized mice had high levels of replication-competent virus in the lip indicating productive infection; however, replication-competent virus was not detected in the lip from any immunized mouse (Figure 19A). By qPCR, the PBS mice had an average 5.8 log10 of HSV-1 DNA present. All vaccinated mice had at least a 1.6-fold reduction in HSV-1 DNA copies. The 10µg tri-HSV-2 modified RNA vaccine immunized group had a significantly lower copy number of viral DNA compared to the 10µg tri-HSV-1 modified RNA vaccine immunized group (Figure 19B). Infectious virus in the TG was measured 5 days post-challenge. Five of 10 (50%) PBS mice had replication competent virus present in the TG, while no immunized mouse had replication competent virus (Figure 19C). The presence of replication competent virus or HSV-1 DNA in the TG indicates a risk of virus establishing a latent infection. Four of 10 (40%) of the PBS mice had viral DNA in the TG at 5 days (Figure 19D). Only 1/40 (2.5%) tri-HSV-1 or tri-HSV-2 modified RNA vaccine immunized mice was positive for viral DNA present in the TG at 5 days. The single positive animal was in the 1-μg group of tri-HSV-1 modified RNA vaccine immunized mice (Figure 19D). [0616] Mice were monitored for weight loss and lip lesions over 13 days. The PBS vaccinated mice lost approximately 5% body weight between days 6 and 9 post-infection. After day 10, the weight in the PBS mice recovered (Figure 19E). None of the immunized mice lost more than 3% body weight (Figure 19E). The PBS mice developed lesions on the lower lip 5 days post-infection. At 6 days, 22/24 (92%) of PBS mice had lower lip lesions. The lesions healed over the next 4 days and were no longer present by day 11 (Figure 19F). None of the immunized mice developed lip lesions (Figure 19F). The TGs were analyzed for HSV-1 DNA copy number 28 days post- challenge as an indicator of latent HSV-1 infection. HSV-1 DNA in the TG was detected from 79% of the PBS mice (Figure 19G). Only a single vaccinated mouse in the 1 µg dose tri-HSV-2 modified RNA vaccine immunized group had HSV-1 DNA detected in the TG. The mean copy 344 11979815v1 P-627574-PC number of HSV-1 DNA in PBS mice was 2.7 log10 whereas the HSV-1 DNA copy number in the single breakthrough case in the tri-HSV-2 modified RNA vaccine immunized mouse was 1.1 log10, a 2.5-fold reduction. These data suggest that the tri-HSV-1 and tri-HSV-2 modified RNA vaccines provide excellent protection against high-dose HSV-1 challenge in the lip. Both vaccines were nearly perfect in preventing HSV-1 DNA from reaching the TG at acute and latent time points. The tri-HSV-1 and tri-HSV-2 modified RNA vaccines had equivalent efficacy, even at a low dose of 1 µg total RNA. [0617] Previously, it had been observed that 16/29 (55%) mice immunized with tri-HSV-2 modified RNA vaccine had positive day 2 vaginal virus titers (mean titer 1.3 PFU/mL) after HSV- 1 intravaginal challenge. An evaluation of whether the tri-HSV-1 modified RNA vaccine was better than the tri-HSV-2 modified RNA vaccine in reducing day 2 or day 4 vaginal virus titers after HSV-1 challenge was performed. Mice were vaccinated with tri-HSV-1 and tri-HSV-2 modified RNA vaccines at low (1 µg) and high (10 µg) doses, treated with Depo-Provera five days before infection, and challenged with 2×106 PFU of HSV-1 strain NS. The PBS immunized animals started succumbing to infection by day 8. By day 14, only 1 of 5 (20%) survived the HSV- 1 challenge, while none of the tri-HSV-1 or tri-HSV-2 modified RNA vaccinated mice died, at either1 µg or 10 µg doses (Figure 20A). In the PBS group, mice started to lose weight by 6 days post-challenge, while the tri-HSV-1 and tri-HSV-2 modified RNA vaccine immunized mice had little or no weight loss (Figure 20B). Mice were scored for genital disease over 14 days. The PBS vaccinated mice developed genital disease on day 5 post-challenge that peaked on day 11 with a mean disease score of 2.8. Genital disease was not observed in any vaccinated mouse (Figure 20C). [0618] An evaluation of whether infectious virus was present in vaginal swabs obtained on days 2 or 4 post-challenge was performed. High titers of HSV-1 were detected in all the PBS animals, with a mean titer of 106 PFU/mL. Virus titers were significantly lower in all vaccine groups on days 2 (Figure 20D) and 4 (Figure 20E). Importantly, day 2 virus titers for the 1 µg tri-HSV-1 modified RNA vaccine immunized group were approximately 2 log10 lower than the 1 µg tri-HSV- 2 modified RNA vaccine immunized group. This difference was the only statistically significant difference detected favoring the tri-HSV-1 modified RNA vaccine in this study and was limited to the 1 µg dose, while at the 10 µg dose in the lip model, the opposite result was observed (Figure 19B), i.e., there was a lower HSV-1 DNA copy number in the vaginas of tri-HSV-2 modified RNA vaccine immunized mice than tri-HSV-1 modified RAN vaccine immunized mice (Figures 19B 345 11979815v1 P-627574-PC and 20D). Significant reductions in viral titers were observed in all vaccine groups compared to PBS at 4 days post-challenge with no difference between the tri-HSV-1 and tri-HSV2 modified RNA vaccine immunized groups (Figure 20E). HSV-1 DNA copy number present in the DRG at the time of humane sacrifice (PBS group) or 28 days post-challenge was evaluated. High levels of HSV-1 DNA were detected in the four PBS animals that were humanely euthanized, while the one surviving animal did not have HSV-1 DNA detected in the DRG at the end of the experiment (Figure 20F). No significant differences were found between the tri-HSV-1 and tri-HSV-2 modified RNA vaccine immunized groups. These results demonstrate that both the tri-HSV-1 and tri-HSV-2 modified RNA vaccines protected mice against a high dose HSV-1 intravaginal challenge, with only a very small difference favoring the tri-HSV-1 modified RNA vaccine immunized group for day 2 vaginal virus titers at the 1 µg dose. Discussion [0619] An effective prophylactic vaccine is the best approach to limit new genital and non-genital HSV infections. The gC2, gD2, and gE2 amino acid sequences share 65%, 82%, and 73% identity with gC1, gD1, and gE1, respectively. An evaluation was performed to determine whether this level of identity was sufficient to generate immune responses capable of potent cross-protection against HSV-1 infection at non-genital sites in mice and whether the protection provided by a tri- HSV-1 vaccine against genital HSV-1 infection in mice can be improved. [0620] The most common site of HSV-1 infection is orolabial. In contrast, HSV-2 rarely infects the oral cavity, suggesting that HSV-1 has a fitness advantage at this site. The lip scarification model of HSV-1 infection was chosen to compare immunization efficacy of the exemplary tri- HSV-2 modified RNA vaccine with an exemplary tri-HSV-1 modified RNA vaccine. In this model, the epithelial surface is scratched to allow the virus to infect the underlying susceptible cells. The tri-HSV-2 RNA vaccine immunization provided total protection against clinical and subclinical infection. Importantly, tri-HSV-2 modified RNA vaccine immunized mice were completely protected against HSV-1 DNA reaching the TG, indicating a minimal risk of latency and reactivation. [0621] An assessment was performed to determine whether the tri-HSV-1 modified RNA vaccine provides better protection than the tri-HSV-2 modified RNA vaccine in the murine genital infection model, particularly against subclinical infection measured by day 2 and day 4 vaginal virus titers. A difference in vaginal titers was observed when comparing the 1 ^g dose of tri-HSV- 346 11979815v1 P-627574-PC 1 modified RNA vaccine with 1 ^g tri-HSV-2 modified RNA vaccine on day 2, but no other differences were noted. [0622] In mice, T-cells play a vital role in restricting the ability of HSV-1 or HSV-2 to reactivate from latency. Mice immunized with tri-HSV-2 modified RNA were shown to have robust CD8+ T-cell responses to gE1 overlapping peptides. Surprisingly, the responses generated by tri-HSV-2 modified RNA vaccine immunized mice were stronger than by tri-HSV-1 modified RNA vaccine immunized mice as manifest by an increase of polyfunctional IFNγ+TNFα+IL-2+ CD8+ T-cells and an increase in CD107a+CD8+ T-cells. A greater CD4+ T cell response to gD1 overlapping peptides was detected in tri-HSV-2 modified RNA vaccine immunized mice than tri-HSV-1 modified RNA vaccine immunized mice. These T cell responses may translate into the tri-HSV-2 modified RNA vaccine having an advantage over the tri-HSV-1 modified RNA vaccine in preventing HSV-1 reactivation from latency. This advantage may be more apparent in the guinea pig genital infection model than the mouse model because recurrent genital lesions and recurrent shedding of HSV DNA can be detected in the guinea pig model. Although antibody responses to gC1 and gE1 were lower for the tri-HSV-2 modified RNA vaccine than the tri-HSV-1 modified RNA vaccine, the T- cell responses were more robust. The potent CD8+ T cell responses reveal an unexpected benefit of the tri-HSV-2 modified RNA vaccine against HSV-1 infection. EXAMPLE 19: EVALUATION OF THE HSV-2 TRIVALENT NUCLEOSIDE- MODIFIED RNA/LNP VACCINE AGAINST HSV-1 ENCEPHALITIS IN MICE [0623] HSV-1 is the most common cause of sporadic infectious encephalitis worldwide and carries approximately a 30% mortality and 60% to 70% morbidity in survivors despite treatment with acyclovir. A vaccine that prevents HSV-1 encephalitis would represent a major advance. [0624] Female BALB/c mice at age 6-8 weeks were immunized intramuscularly in a hind limb twice at one-month intervals with PBS (control), with 1 ^g trivalent HSV-2 RNA vaccine containing 0.33 ^g each of exemplary gC2, gD2, gE2 nucleoside-modified RNA in lipid nanoparticle (LNP), or with 10 ^g trivalent HSV-2 RNA vaccine containing 3.33 ^g each of exemplary gC2, gD2, gE2 nucleoside-modified RNA in LNP. One month after the second immunization, mice were infected intranasally with 5x105 PFU of HSV-1 (100 LD50) and evaluated for survival, neurologic disease, and for HSV-1 DNA copy number by qPCR in brain, trigeminal ganglia, and olfactory bulb tissues harvested post-infection. [0625] Survival. Naïve (not vaccinated) 6 to 8-week-old female mice were inoculated intranasally with 5x103, 5x104, 5x105, 5x106 plaque-forming units (pfu) of HSV-1 strain H129, or with PBS 347 11979815v1 P-627574-PC as a control. Survival was evaluated over 28 days. All mice in the PBS group survived, while all mice in the groups inoculated with 5x104, 5x105, or 5x106 pfu required humane euthanasia (Figure 21). Survival was 50% (2 of 4 mice) at 5x103 pfu, establishing the LD50 as 5x103 pfu (Figure 21). [0626] Trivalent HSV-2 vaccine protects BALB/c mice against HSV-1 encephalitis. All 10 animals in the vaccine group immunized with 1 ^g or 10 ^g of the tri-HSV-2 RNA-LNP survived, while all 10 mice in the PBS group required humane euthanasia on day 6 post-infection (Figure 22A). HSV-1 DNA was detected in brains in 5 of 5 (100%) mice in the PBS group with a mean titer of 7 log10 HSV-1 DNA copies per 105 adipsin genes, while HSV-1 DNA was detected in 2 of 10 (20%) mice in the 1 ^g group with a mean titer of 0.4 log10, or 1 of 10 (10%) mice in the 10 ^g group with a mean titer of 0.2 log10 (Figure 22B). HSV-1 DNA was detected in the trigeminal ganglia in 5 of 5 (100%) mice from the PBS group at a mean titer of 6.7 log10, while HSV-1 DNA was detected in 9 of 10 (90%) mice in the 1 ^g group with a mean titer of 2.8 log10, or 7 of 10 (70%) mice in the 10 ^g group with a mean titer of 2.0 log10 (Figure 22C). HSV-1 DNA was detected in the olfactory bulb of mice in the PBS group in 5 of 5 (100%) mice at a mean titer of 7.2 log10, while HSV-1 DNA was detected in 2 of 9 (22%) mice in the 1 ^g group with a mean titer of 0.6 log10, or 2 of 10 (20%) mice in the 10 ^g group with a mean titer of 0.4 log10 (Figure 22D). [0627] Conclusions: The 1 and 10 ^g doses of the trivalent HSV-2 vaccine were both highly efficacious at protecting mice against death and preventing HSV-1 DNA from reaching the brain and the olfactory bulb. The vaccine was less effective at preventing HSV-1 DNA from reaching the trigeminal ganglia, although the vaccine groups had significantly reduced HSV-1 DNA copy number in the trigeminal ganglia compared with the PBS group. The HSV-2 nucleoside-modified RNA-LNP vaccine is highly efficacious at preventing HSV-1 encephalitis in a mouse model following a high dose lethal challenge. [0628] Having described preferred embodiments of the invention with reference to the accompanying drawings, it is to be understood that the invention is not limited to the precise embodiments, and that various changes and modifications may be effected therein by those skilled in the art without departing from the scope or spirit of the invention as defined in the appended claims. [0629] All patent documents and references cited herein are incorporated by reference as if fully set forth. 348 11979815v1

Claims

P-627574-PC CLAIMS What is claimed is: 1. A method of treating or inhibiting a herpes simplex virus (HSV) oral mucosal infection, an HSV encephalitis infection, or a combination thereof in a subject comprising administering to said subject: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof. 2. The method of claim 1, wherein said infection is a primary infection or a secondary infection. 3. The method of claim 1 or claim 2, wherein one or more of said RNAs comprise one or more ribonucleic acids that are nucleoside modified. 4. The method of claim 3, wherein the one or more nucleoside modified ribonucleic acids comprise one or more m5D (5-methyldihydrouridine) residues. 5. The method of claim 3, wherein the one or more nucleoside modified ribonucleic acids comprise one or more pseudouridine residues. 6. The method of claim 5, wherein said one or more pseudouridine residues comprise m1Ψ (1- methylpseudouridine). 7. The method of claim 6, wherein said one or more pseudouridine residues comprise m1acp3Ψ (1-methyl-3-(3-amino-5-carboxypropyl)pseudouridine, Ψm (2′-O-methylpseudouridine, m3Ψ (3-methylpseudouridine), or any combination thereof. 8. The method of any one of claims 1-7, wherein: (a) the HSV gC or an immunogenic fragment thereof is an HSV-2 gC or immunogenic fragment thereof; (b) the HSV gD or an immunogenic fragment thereof is an HSV-2 gD or immunogenic fragment thereof; (c) the HSV gE or an immunogenic fragment thereof is an HSV-2 gE or immunogenic fragment thereof; or (d) any combination thereof. 349 11979815v1 P-627574-PC 9. The method of claim 8, wherein: (a) the HSV-2 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 28, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 241, or SEQ ID NO: 242; (b) the HSV-2 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 110, SEQ ID NO: 234, or SEQ ID NO: 120; (c) the HSV-2 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 17 or SEQ ID NO: 18; or (d) any combination thereof. 10. The method of claim 8, wherein: (a) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 25, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239, or SEQ ID NO: 246; (b) the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 23, SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, or SEQ ID NO: 247; (c) the RNA encoding HSV-2 gE or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 27, SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, or SEQ ID NO: 148; or (d) any combination thereof. 11. The method of claim 8, wherein: (a) the HSV-2 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 224; (b) the HSV-2 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 5; 350 11979815v1 P-627574-PC (c) the HSV-2 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 17; or (d) any combination thereof. 12. The method of claim 8, wherein: (a) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 246; (b) the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 247; (c) the RNA encoding HSV-2 gE or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 145; or (d) any combination thereof. 13. The method of claim 12, wherein: (a) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 246; (b) the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 247; and (c) the RNA encoding HSV-2 gE or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 145. 14. The method of any one of claims 1-7, wherein: (a) the HSV gC or an immunogenic fragment thereof is an HSV-1 gC or immunogenic fragment thereof; (b) the HSV gD or an immunogenic fragment thereof is an HSV-1 gD or immunogenic fragment thereof; (c) the HSV gE or an immunogenic fragment thereof is an HSV-1 gE or immunogenic fragment thereof; or (d) any combination thereof. 15. The method of claim 14, wherein: (a) the HSV-1 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 240; (b) the HSV-1 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 134; 351 11979815v1 P-627574-PC (c) the HSV-1 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 248, or SEQ ID NO: 140; or (d) any combination thereof. 16. The method of claim 14, wherein: (a) the RNA encoding HSV-1 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 24; (b) the RNA encoding HSV-1 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 22; (c) the RNA encoding HSV-1 gE or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 26; or (d) any combination thereof. 17. The method of any one of claims 1-16, wherein: (a) the immunogenic fragment of HSV gC comprises the ectodomain of HSV gC, (a) the immunogenic fragment of HSV gD comprises the ectodomain of HSV gD, (b) the immunogenic fragment of HSV gE comprises the ectodomain of HSV gE, or (c) any combination thereof. 18. The method of any one of claims 1-17, further comprising administering to the subject one or more RNAs encoding (a) HSV glycoprotein B (gB) or immunogenic fragment thereof, (b) HSV glycoprotein H (gH) or immunogenic fragment thereof, (c) HSV glycoprotein L (gL) or immunogenic fragment thereof, (d) HSV glycoprotein I (gI) or immunogenic fragment thereof, or (e) any combination thereof. 19. The method of any one of claims 1-18, wherein one or more of said RNAs further comprise a signal sequence encoding a signal peptide. 20. The method of claim 19, wherein the signal peptide comprises an amino acid sequence of any one of SEQ ID NO: 29-48 and 220-222. 21. The method of claim 19, wherein the signal sequence is or comprises any signal sequence listed in Table 4. 22. The method of claim 20, wherein the signal peptide comprises an IL-2 signal peptide. 23. The method of claim 20, wherein the signal peptide comprises a viral signal peptide. 352 11979815v1 P-627574-PC 24. The method of claim 23, wherein the viral signal peptide comprises an HSV signal peptide. 25. The method of claim 24, wherein the HSV signal peptide comprises an HSV gD signal peptide. 26. The method of claim 25, wherein the HSV gD signal peptide comprises an HSV-1 gD signal peptide. 27. The method of claim 26, wherein the HSV-1 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 34 or SEQ ID NO: 35. 28. The method of claim 26, wherein the HSV-1 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 35. 29. The method of claim 25, wherein the HSV gD signal peptide is an HSV-2 gD signal peptide. 30. The method of claim 29, wherein the HSV-2 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, or SEQ ID NO: 33. 31. The method of claim 30, wherein the HSV-2 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 29 or SEQ ID NO: 31. 32. The method of any one of claims 19-31, wherein the signal sequence is a codon optimized signal sequence. 33. The method of any one of claims 1-32, wherein one or more of said RNAs further comprise a poly-A tail. 34. The method of any one of claims 1-33, wherein one or more of said RNAs further comprise a cap. 35. The method of claim 34, wherein the cap is an m7GpppG cap, 3′-O-methyl-m7GpppG cap, or anti-reverse cap analog. 36. The method of any one of claims 1-35, wherein one or more of said RNAs further comprise a cap-independent translational enhancer. 37. The method of any one of claims 1-36, wherein one or more of said RNAs further comprise 5′ and/or 3′ untranslated regions. 38. The method of any one of claims 1-37, wherein one or more of said RNAs are formulated with a nanoparticle, lipid, polymer, cholesterol, cell penetrating peptide, or any combination thereof. 39. The method of claim 38, wherein said nanoparticle is a lipid nanoparticle. 353 11979815v1 P-627574-PC 40. The method of any one of claims 1-39, wherein administration comprises intramuscular administration. 41. The method of any one of claims 1-39, wherein administration comprises subcutaneous administration. 42. The method of any one of claims 1-39, wherein administration comprises intradermal administration. 43. The method of any one of claims 1-39, wherein administration comprises intranasal, intravaginal, or intrarectal administration. 44. The method of any one of claims 1-39, wherein administration comprises topical administration. 45. The method of any one of claims 1-44, wherein the first composition, second composition, and third composition are different compositions. 46. The method of any one of claims 1-44, wherein the first composition, second composition, and third composition are the same composition. 47. The method of any one of claims 1-46, wherein administering to said subject comprises administering one or more doses of each of the first composition, second composition, or third composition. 48. The method of any one of claims 1-47, wherein said HSV oral mucosal infection, HSV encephalitis infection, or both are HSV-1 infections. 49. A method of preventing establishment of a latent Herpes Simplex Virus (HSV) infection in a subject comprising administering to said subject: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof. 50. The method of claim 49, wherein one or more of said RNAs comprise one or more ribonucleic acids that are nucleoside modified. 51. The method of claim 49 or claim 50, wherein the one or more nucleoside modified ribonucleic acids comprise one or more m5D (5-methyldihydrouridine) residues. 354 11979815v1 P-627574-PC 52. The method of claim 49 or claim 50, wherein the one or more nucleoside modified ribonucleic acids comprise one or more pseudouridine residues. 53. The method of claim 52, wherein said one or more pseudouridine residues comprise m1Ψ (1-methylpseudouridine). 54. The method of claim 52, wherein said one or more pseudouridine residues comprise m1acp3Ψ (1-methyl-3-(3-amino-5-carboxypropyl)pseudouridine, Ψm (2′-O- methylpseudouridine, m3Ψ (3-methylpseudouridine), or any combination thereof. 55. The method of any one of claims 49-54, wherein: (a) the HSV gC or an immunogenic fragment thereof is an HSV-2 gC or immunogenic fragment thereof, (b) the HSV gD or an immunogenic fragment thereof is an HSV-2 gD or immunogenic fragment thereof, (c) the HSV gE or an immunogenic fragment thereof is an HSV-2 gE or immunogenic fragment thereof, (d) any combination thereof. 56. The method of claim 55, wherein: (a) the HSV-2 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 28, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 241, or SEQ ID NO: 242; (b) the HSV-2 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 110, SEQ ID NO: 234, or SEQ ID NO: 120; (c) the HSV-2 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 17 or SEQ ID NO: 18; or (d) any combination thereof. 57. The method of claim 55, wherein: (a) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 25, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 218, SEQ 355 11979815v1 P-627574-PC ID NO: 219, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 239, or SEQ ID NO: 246; (b) the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 23, SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, or SEQ ID NO: 247; (c) the RNA encoding HSV-2 gE or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 27, SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, or SEQ ID NO: 148; or (d) any combination thereof. 58. The method of claim 56, wherein: (a) the HSV-2 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 224; (b) the HSV-2 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 5; (c) the HSV-2 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 17; or (d) any combination thereof. 59. The method of claim 57, wherein: (a) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 246; (b) the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 247; (c) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 145; or (d) any combination thereof. 60. The method of claim 59, wherein: (a) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 246; (b) the RNA encoding HSV-2 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 247; and 356 11979815v1 P-627574-PC (c) the RNA encoding HSV-2 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 145. 61. The method of any one of claims 49-60, wherein: (a) the HSV gC or an immunogenic fragment thereof is an HSV-1 gC or immunogenic fragment thereof, (b) the HSV gD or an immunogenic fragment thereof is an HSV-1 gD or immunogenic fragment thereof, (c) the HSV gE or an immunogenic fragment thereof is an HSV-1 gE or immunogenic fragment thereof, or (d) any combination thereof. 62. The method of claim 61, wherein: (a) the HSV-1 gC or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 240; (b) the HSV-1 gD or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 134; (c) the HSV-1 gE or immunogenic fragment thereof comprises an amino acid sequence at least 85% identical to SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 248, or SEQ ID NO: 140; or (d) any combinations thereof. 63. The method of claim 61, wherein: (a) the RNA encoding HSV-1 gC or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 24; (b) the RNA encoding HSV-1 gD or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 22; (c) the RNA encoding HSV-1 gE or an immunogenic fragment thereof comprises a nucleotide sequence at least 85% identical to SEQ ID NO: 26; or (d) any combinations thereof. 64. The method of claim 61, wherein: (a) the immunogenic fragment of HSV gC comprises the ectodomain of HSV gC, (b) the immunogenic fragment of HSV gD comprises the ectodomain of HSV gD, (c) the immunogenic fragment of HSV gE comprises the ectodomain of HSV gE, or (d) any combination thereof. 357 11979815v1 P-627574-PC 65. The method of any one of claims 49-64, further comprising administering one or more RNAs encoding (a) an HSV glycoprotein B (gB) or immunogenic fragment thereof, (b) an HSV glycoprotein H (gH) or immunogenic fragment thereof, (c) an HSV glycoprotein L (gL) or immunogenic fragment thereof, (d) an HSV glycoprotein I (gI) or immunogenic fragment thereof, or (e) any combination thereof. 66. The method of any one of claims 49-65, wherein one or more of said RNAs further comprise a signal sequence encoding a signal peptide. 67. The method of claim 66, wherein the signal peptide comprises an amino acid sequence of any one of SEQ ID NO: 29-48 and 220-222. 68. The method of claim 66, wherein the signal sequence is or comprises any signal sequence listed in Table 4. 69. The method of claim 66, wherein the signal peptide is an IL-2 signal peptide. 70. The method of claim 66, wherein the signal peptide is a viral signal peptide. 71. The method of claim 70, wherein the viral signal peptide is an HSV signal peptide. 72. The method of claim 71, wherein the HSV signal peptide is an HSV gD signal peptide. 73. The method of claim 72, wherein the HSV gD signal peptide is an HSV-1 gD signal peptide. 74. The method of claim 73, wherein the HSV-1 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 34 or SEQ ID NO: 35. 75. The method of claim 74, wherein the HSV-1 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 35. 76. The method of claim 72, wherein the HSV gD signal peptide is an HSV-2 gD signal peptide. 77. The method of claim 76, wherein the HSV-2 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, or SEQ ID NO: 33. 78. The method of claim 76, wherein the HSV-2 gD signal peptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 29 or SEQ ID NO: 31. 79. The method of any one of claims 66-78, wherein the signal sequence is a codon optimized signal sequence. 80. The method of any one of claims 49-79, wherein one or more of said RNAs further comprise a poly-A tail. 358 11979815v1 P-627574-PC 81. The method of any one of claims 49-80, wherein one or more of said RNAs further comprise a cap. 82. The method of claim 81, wherein the cap is an m7GpppG cap, 3′-O-methyl-m7GpppG cap, or anti-reverse cap analog. 83. The method of any one of claims 49-82, wherein one or more of said RNAs further comprise a cap-independent translational enhancer. 84. The method of any one of claims 49-83, wherein one or more of said RNAs further comprise 5′ and/or 3′ untranslated regions. 85. The method of any one of claims 49-84, wherein one or more of said RNAs are formulated with a nanoparticle, lipid, polymer, cholesterol, cell penetrating peptide, or any combination thereof. 86. The method of claim 85, wherein said nanoparticle is a lipid nanoparticle. 87. The method of any one of claims 49-86, wherein said latent HSV infection comprises an HSV-1 infection. 88. The method of any one of claims 49-86, wherein said latent HSV infection comprises an HSV-2 infection. 89. The method of any one of claims 49-88, wherein administration comprises intramuscular administration. 90. The method of any one of claims 49-88, wherein administration comprises subcutaneous administration. 91. The method of any one of claims 49-88, wherein administration comprises intradermal administration. 92. The method of any one of claims 49-88, wherein administration comprises intranasal, intravaginal, or intrarectal administration. 93. The method of any one of claims 49-88, wherein administration comprises topical administration. 94. The method of any one of claims 49-93, wherein the first composition, second composition, and third composition are different compositions. 95. The method of any one of claims 49-93, wherein the first composition, second composition, and third composition are the same composition. 359 11979815v1 P-627574-PC 96. The method of any one of claims 49-95, wherein administration to said subject comprises administering one or more doses of each of the first composition, second composition, or third composition. 97. The method of any one of claims 49-96, wherein said latent HSV infection is in the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or any combination thereof. 98. A method of treating or inhibiting a Herpes Simplex Virus (HSV) infection of the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or a combination thereof in a subject comprising administering to said subject: (a) a first composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a second composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a third composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof. 99. The method of claim 98, wherein said HSV infection is an HSV-1 infection. 100. A combination for use in treating or inhibiting a Herpes Simplex Virus (HSV) oral mucosal infection, an HSV encephalitis infection, or a combination thereof; or an HSV-1 infection, wherein the combination comprises: (a) a composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions. 101. The combination of claim 100, wherein the infection is a latent infection, a primary infection, or a secondary infection. 102. The combination of claim 100, wherein the infection is in the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or any combinations thereof. 360 11979815v1 P-627574-PC 103. Use of a combination for treating or inhibiting Herpes Simplex Virus (HSV) oral mucosal infection, encephalitis infection, or a combination thereof; or an HSV-1 infection, wherein the combination comprises: (a) a composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions. 104. Use of a combination for the manufacture of a medicament for treating or inhibiting Herpes Simplex Virus (HSV) oral mucosal infection, an HSV encephalitis infection, or a combination thereof; or an HSV-1 infection, wherein the combination comprises: (a) a composition comprising RNA encoding HSV glycoprotein D (gD) or an immunogenic fragment thereof, (b) a composition comprising RNA encoding HSV glycoprotein C (gC) or an immunogenic fragment thereof, and (c) a composition comprising RNA encoding HSV glycoprotein E (gE) or an immunogenic fragment thereof, wherein the use comprises administering one or more doses of one or more of the compositions. 105. The use of claim 103 or 104, wherein the infection is a latent infection, a primary infection, or a secondary infection. 106. The use of any one of claims 103-105, wherein the infection is in the dorsal root ganglia, trigeminal ganglia, brain, olfactory bulb, or any combination thereof. 361 11979815v1
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WO2019035066A1 (en) * 2017-08-17 2019-02-21 The Trustees Of The University Of Pennsylvania Modified mrna vaccines encoding herpes simplex virus glycoproteins and uses thereof

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WO2019035066A1 (en) * 2017-08-17 2019-02-21 The Trustees Of The University Of Pennsylvania Modified mrna vaccines encoding herpes simplex virus glycoproteins and uses thereof
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