RU2823353C2 - CODON-OPTIMIZED EXPRESSION CASSETTES OF ACID α-GLUCOSIDASE AND METHODS OF USE THEREOF - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology and genetic engineering, in particular, to nucleic acids coding acid alpha-glucosidase (GAA). Present invention also relates to expression cassettes, vectors, cells and cell lines and methods of using such nucleic acids coding acid alpha-glucosidase (GAA).

EFFECT: invention enables to use the invention as a gene therapy of individuals with Pompe disease and other diseases and disorders which can be treated using GAA.

86 cl, 21 dwg, 3 tbl, 14 ex

Description

Cross reference to related applications

[0001] This application claims benefit from U.S. Provisional Patent Application No. 62/672419, filed May 16, 2018, and U.S. Provisional Patent Application No. 62/734454, filed September 21, 2018. The entire contents of the above applications are incorporated herein by reference, including all text, tables, sequence listings, and drawings.

Introduction

[0002] Glycogenosis type II, also known as Pompe disease, is an autosomal recessive disorder caused by mutations in the gene encoding the lysosomal enzyme α-glucosidase (GAA), which catalyzes glycogen degradation. The resulting enzyme deficiency leads to pathological glycogen accumulation and lysosomal changes in all tissues of the body, leading to cardiac, respiratory and skeletal muscle dysfunction (van der Ploeg & Reuser, 2008 Lancet, 372:1342-1353). Pompe disease is generally divided into two main phenotypes—infantile form of Pompe disease (IOPD) (also referred to as infantile form of Pompe disease (IPD) or early-onset Pompe disease) and late-onset Pompe disease (LOPD)—based on residual GAA enzyme activity , age of onset, organ involvement (i.e. presence of cardiomyopathy), severity and rate of progression. For Pompe disease, replacement therapy (ERT) is available; however, it has some limitations (i.e. limited biodistribution and high immunogenicity), leading to treatment failure and limited long-term effectiveness (van der Ploeg & Reuser, 2008).

The essence of the invention

[0003] The present invention relates to optimized cassettes for liver-directed expression of a secreted version of human GAA. This cassette optimization results in increased hepatic GAA secretion and enables hepatic gene transfer to achieve circulating levels of GAA sufficient to systemically correct GAA deficiency in individuals. These cassettes achieve increased transgene expression, improved safety characteristics and potentially reduced immunogenicity. These cassettes could be used as gene therapy for individuals with Pompe disease and other diseases and disorders that can be treated using GAAs.

[0004] Codon optimization of the GAA expression cassette was performed to improve GAA expression. In one embodiment, the nucleic acid sequences encoding the GAAs have been modified to eliminate CpG dinucleotides. A total of 20 new codon-optimized transgene sequences (GAA1-GAA20) were generated. Given the comparison of in vitro GAA activity, 5 codon-optimized sequences (GAA 2, 5, 7, 8 and 13) were selected for further optimization. The GAA13 sequence was used to analyze differences in GAA expression after adding a 29-bp polynucleotide sequence to the 5′ untranslated region (UTR). For GAA7, 8 and 13, two different polyadenylation sequences derived from bovine growth hormone (bGH or BGH) (wild type and CpG reduced) were also evaluated. The resulting 9 expression cassettes (Table 1) were packaged into a capsid SEQ ID NO: 30-32; one was also packaged into the AAV6 capsid.

[0005] The present invention relates to nucleic acids encoding acid α-glucosidase (GAA), expression cassettes containing nucleic acids encoding acid α-glucosidase (GAA), and viral vectors containing nucleic acids encoding acid α-glucosidase (GAA ).

[0006] In one embodiment, the nucleic acid encoding a GAA has greater than 86% sequence identity to any of the sequences shown as SEQ ID NO: 1-5. In additional aspects, the GAA-encoding nucleic acid has greater than 87% sequence identity to any of the sequences set forth in SEQ ID NOs: 1-5.

[0007] In another embodiment, the nucleic acid encoding a GAA has greater than 87% sequence identity to any of the sequences shown as SEQ ID NO: 6-15. In particular aspects, the nucleic acid encoding a GAA has greater than 88% sequence identity to any of the sequences set forth in SEQ ID NO: 6-15. In further specific aspects, the GAA-encoding nucleic acid has greater than 89% sequence identity to any of the sequences set forth in SEQ ID NOs: 6-15. In further specific aspects, the nucleic acid encoding a GAA has greater than 90% sequence identity to any of the sequences set forth in SEQ ID NO: 6-15. In further specific aspects, the GAA-encoding nucleic acid has greater than 91% sequence identity to any of the sequences set forth in SEQ ID NO: 6-15.

[0008] In another embodiment, the nucleic acid encoding a GAA has greater than 91% sequence identity to any of the sequences shown as SEQ ID NO: 16-24.

[0009] In another embodiment, the nucleic acid encoding a GAA has greater than 92% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0010] In another embodiment, the nucleic acid encoding a GAA has greater than 93% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0011] In another embodiment, the nucleic acid encoding a GAA has greater than 94% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0012] In another embodiment, the nucleic acid encoding a GAA has greater than 95% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0013] In another embodiment, the nucleic acid encoding a GAA has greater than 96% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0014] In another embodiment, the nucleic acid encoding a GAA has greater than 97% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0015] In another embodiment, the nucleic acid encoding a GAA has greater than 98% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0016] In another embodiment, the nucleic acid encoding a GAA has greater than 99% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0017] In another embodiment, the nucleic acid encoding a GAA has greater than 99.5% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0018] In another embodiment, the nucleic acid encoding a GAA has 100% sequence identity to any of the sequences shown as SEQ ID NO: 1-24.

[0019] In another embodiment, the nucleic acid encoding a GAA contains less than 127 CpG dinucleotides.

[0020] In another embodiment, the nucleic acid encoding a GAA contains less than 126 CpG dinucleotides.

[0021] In another embodiment, the nucleic acid encoding a GAA contains approximately 126-120 CpG dinucleotides.

[0022] In another embodiment, the nucleic acid encoding a GAA contains approximately 120-110 CpG dinucleotides.

[0023] In another embodiment, the nucleic acid encoding a GAA contains approximately 110-100 CpG dinucleotides.

[0024] In another embodiment, the nucleic acid encoding a GAA contains approximately 100-90 CpG dinucleotides.

[0025] In another embodiment, the nucleic acid encoding a GAA contains approximately 90-80 CpG dinucleotides.

[0026] In another embodiment, the nucleic acid encoding a GAA contains approximately 80-70 CpG dinucleotides.

[0027] In another embodiment, the nucleic acid encoding a GAA contains approximately 70-60 CpG dinucleotides.

[0028] In another embodiment, the nucleic acid encoding a GAA contains approximately 60-50 CpG dinucleotides.

[0029] In another embodiment, the nucleic acid encoding a GAA contains approximately 50-40 CpG dinucleotides.

[0030] In another embodiment, the nucleic acid encoding a GAA contains approximately 40-30 CpG dinucleotides.

[0031] In another embodiment, the nucleic acid encoding a GAA contains approximately 30-20 CpG dinucleotides.

[0032] In another embodiment, the nucleic acid encoding a GAA contains less than 20 CpG dinucleotides.

[0033] In another embodiment, the nucleic acid encoding a GAA contains approximately 20-10 CpG dinucleotides.

[0034] In another embodiment, the nucleic acid encoding a GAA contains less than 10 CpG dinucleotides.

[0035] In another embodiment, the nucleic acid encoding a GAA contains approximately 10-5 CpG dinucleotides.

[0036] In another embodiment, the nucleic acid encoding a GAA contains 5 CpG dinucleotides.

[0037] In another embodiment, the nucleic acid encoding a GAA contains 4 CpG dinucleotides.

[0038] In another embodiment, the nucleic acid encoding a GAA contains 3 CpG dinucleotides.

[0039] In another embodiment, the nucleic acid encoding a GAA contains 2 CpG dinucleotides.

[0040] In another embodiment, the nucleic acid encoding a GAA contains 1 CpG dinucleotide.

[0041] In another embodiment, the nucleic acid encoding a GAA contains 0 CpG dinucleotides.

[0042] The invention also provides expression cassettes containing nucleic acids encoding a GAA, as defined herein, operably linked to an expression control element.

[0043] In one embodiment, the expression cassette contains a nucleic acid encoding a GAA with greater than 86% sequence identity (eg, 87-100% identity) to any of the sequences shown as SEQ ID NO: 1-5.

[0044] In another embodiment, the expression cassette contains a nucleic acid encoding a GAA with greater than 87% sequence identity (eg, 88%-100% identity) to any of the sequences shown as SEQ ID NO: 6-15.

[0045] In another embodiment, the expression cassette contains a nucleic acid encoding a GAA with less than 127 CpG dinucleotides, less than 127 CpG dinucleotides, less than 127 CpG dinucleotides, less than 126 CpG dinucleotides, less than 125 CpG dinucleotides, less than 124 CpG dinucleotides, less than 123 CpG dinucleotides, less than 122 CpG dinucleotides, less than 121 CpG dinucleotides, less than 120 CpG dinucleotides, less than 119 CpG dinucleotides, less than 118 CpG dinucleotides, less than 117 CpG dinucleotides, less than 116 CpG dinucleotides , less than 115 CpG dinucleotides, less than 114 CpG dinucleotides, less than 113 CpG dinucleotides, less than 112 CpG dinucleotides, less than 111 CpG dinucleotides, less than 110 CpG dinucleotides, less than 109 CpG dinucleotides, less than 108 CpG dinucleotides, less less than 107 CpG dinucleotides, less than 106 CpG dinucleotides, less than 105 CpG dinucleotides, less than 104 CpG dinucleotides, less than 103 CpG dinucleotides, less than 102 CpG dinucleotides, less than 101 CpG dinucleotides, less than 101 CpG dinucleotides, etc. . to zero (0) CpG dinucleotides.

[0046] In one embodiment, the expression control element is located in the 5' direction relative to the nucleic acid encoding the GAA.

[0047] In another embodiment, the expression cassette includes a polyadenylation (poly-A) sequence located 3' to the nucleic acid encoding the GAA.

[0048] In another embodiment, the expression control element or polyadenylation sequence has a reduced CpG content compared to the wild-type expression control element or polyadenylation sequence.

[0049] In another embodiment, the expression control element comprises an ApoE/hAAT enhancer/promoter sequence.

[0050] In another embodiment, the polyadenylation sequence comprises a bovine growth hormone (bGH) polyadenylation sequence.

[0051] In another embodiment, the ApoE/hAAT enhancer/promoter sequence or bGH polyadenylation sequence has reduced CpG content compared to the ApoE/hAAT enhancer/promoter sequence or wild-type bGH polyadenylation sequence.

[0052] In one aspect, the wild-type bGH polyadenylation sequence comprises the sequence SEQ ID NO: 27.

[0053] In another aspect, the wild-type ApoE/hAAT enhancer/promoter sequence comprises SEQ ID NO: 28 or 29.

[0054] In another aspect, the CpG reduced polyadenylation sequence of bGH comprises the sequence SEQ ID NO: 26.

[0055] In another embodiment, the expression cassette further comprises an intron located between the 3' end of the expression control element and the 5' end of the nucleic acid encoding the GAA.

[0056] In another embodiment, the GAA contains or consists of the sequence shown as SEQ ID NO: 25.

[0057] The invention further relates to viral vectors, such as adeno-associated virus (AAV) vectors, containing nucleic acids encoding GAAs, as defined herein.

[0058] In one embodiment, a viral vector, such as an adeno-associated virus (AAV) vector, contains any of the GAA-encoding nucleic acids as defined herein operably linked to an expression control element.

[0059] In another embodiment, a viral vector, such as an adeno-associated virus (AAV) vector, contains any of the expression cassettes containing GAA-encoding nucleic acids as defined herein.

[0060] In another embodiment, the AAV vector comprises: one or more AAV capsids and one or more AAV inverted terminal repeats (ITRs), wherein the AAV ITRs flank the 5' or 3' end of the nucleic acid or expression cassette.

[0061] In additional embodiments, the AAV vector further comprises an intron located 5' or 3' to one or more ITRs.

[0062] In further embodiments, an AAV vector comprising at least one or more ITRs or introns contains one or more ITRs or introns modified to have reduced CpG content.

[0063] In additional embodiments, the AAV vector has a capsid serotype comprising a modified AAV capsid VP1, VP2 and/or VP3 or a variant thereof having 90% or more sequence identity to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7 , AAV8, AAV9, AAV10, AAV11, Rh10, Rh74, AAV3B, AAV-2i8 or VP1, VP2 and/or VP3 sequences SEQ ID NO: 30, 31 or 32, or a capsid having 95% or more sequence identity to AAV1 , AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, Rh10, Rh74, AAV3B, AAV-2i8 or VP1, VP2 and/or VP3 sequences SEQ ID NO: 30, 31 or 32 , or a capsid having 100% sequence identity to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, Rh10, Rh74, AAV3B, AAV-2i8, or the sequences VP1, VP2 and/ or VP3 SEQ ID NO: 30, 31 or 32.

[0064] In additional embodiments, the AAV vector containing one or more ITRs contains one or more ITRs from any of the AAV serotypes AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, Rh10, Rh74 or AAV3B or combinations thereof.

[0065] The invention further relates to pharmaceutical compositions containing any of nucleic acids encoding GAA, expression cassettes containing nucleic acids encoding GAA, or viral vectors, such as AAV vectors containing nucleic acids encoding GAA, or expression cassettes containing GAA-encoding nucleic acids as defined herein.

[0066] In one embodiment, the pharmaceutical composition contains a plurality of AAV vectors, as defined herein, in a biocompatible carrier or excipient.

[0067] In another embodiment, a pharmaceutical composition comprising any of the AAV vectors as defined herein further comprises empty AAV capsids.

[0068] In specific embodiments, in a pharmaceutical composition comprising AAV vectors and empty AAV capsids, the ratio of empty AAV capsids to AAV vector is in the range of or about 100:1-50:1, about 50:1-25:1, about 25 :1-10:1, approximately 10:1-1:1, approximately 1:1-1:10, approximately 1:10-1:25, approximately 1:25-1:50 or approximately 1:50-1: 100.

[0069] In specific aspects, in a pharmaceutical composition containing AAV vectors and empty AAV capsids, the ratio of empty AAV capsids to AAV vector is approximately 2:1, 3:1, 4:1, 5:1, 6:1, 7: 1, 8:1, 9:1 or 10:1.

[0070] In another embodiment, the pharmaceutical composition includes a surfactant.

[0071] The invention further relates to methods of treating a human in need of α-glucosidase acid (GAA) by administering or delivering to the human any of GAA-encoding nucleic acids, expression cassettes containing GAA-encoding nucleic acids, or viral vectors, such as AAV vectors containing nucleic acids encoding GAA or expression cassettes containing nucleic acids encoding GAA.

[0072] In one embodiment, a method of treating a human in need of α-glucosidase acid (GAA) includes: (a) producing a nucleic acid, expression cassette, or viral vector, such as an AAV vector as defined herein, or any a pharmaceutical composition as defined herein; and (b) administering an amount of nucleic acid, expression cassette, viral (eg, AAV) vector or pharmaceutical composition to a human, wherein the GAA is expressed in the human.

[0073] In specific embodiments, the person has Pompe disease, such as infantile form of Pompe disease or late-onset Pompe disease.

[0074] In another embodiment, the person has glycogen storage disease (GSD).

[0075] In specific aspects, the GSD is any of: GSD type I (Gierke's disease), GSD type III (Forbes disease), GSD type IV (Andersen's disease, amylopectinosis), GSD type V (McArdle's disease), GSD type VI (Hers disease), GSD type VII (Tarui disease) or congenital GSD heart (eg, lethal congenital GSD heart).

[0076] In specific embodiments, a GAA-encoding nucleic acid, an expression cassette containing a GAA-encoding nucleic acid, or an AAV vector is administered to a human intravenously, intraarterially, intracavitarily, intramucosally, or via a catheter.

[0077] In certain embodiments, the method expresses GAA at elevated levels, optionally greater than 1% of the GAA levels found in a GAA-neutral human.

[0078] In specific embodiments, the AAV vector is administered in a range of from about 1×10 ⁸ to about 1×10 ¹⁴ vector genomes per kilogram (KG/kg) of human weight.

[0079] In specific embodiments, the method reduces, reduces, or inhibits one or more symptoms of GAA need or disease; or prevent or reduce the progression or worsening of one or more symptoms of a GAA need or disease; or stabilize one or more symptoms of GAA need or disease; or improve one or more symptoms of a GAA need or disease.

[0080] In specific aspects, a symptom that can be treated by the invention may be one or more of the following: problems eating and/or failure to gain weight; poor head and/or neck control; breathing problems and/or lung infections; enlargement and/or thickening of the heart; heart defects; tongue enlargement; problems with swallowing; liver enlargement; low muscle strength; low muscle tone; weakness in the legs, waist and/or arms; dyspnea; problems with exercise; difficulty breathing during sleep; curvature of the spine and/or ankylosis.

[0081] The invention further relates to cells containing nucleic acids encoding GAA, cells containing expression cassettes containing nucleic acids encoding GAA, and cells containing viral vectors, such as AAV vectors containing nucleic acids encoding GAA or expressing cassettes containing nucleic acids encoding GAA.

[0082] In one embodiment, the cell produces a viral vector.

[0083] In another embodiment, the cell produces an AAV vector, as described herein.

[0084] In addition, the invention also relates to methods for producing viral vectors, such as AAV vectors, as described herein.

[0085] In one embodiment, a method for producing AAV vectors includes: inserting an AAV vector genome containing a nucleic acid encoding a GAA, or an expression cassette containing a nucleic acid encoding a GAA, as defined herein, into a packaging helper cell; and culturing the helper cell under conditions to produce AAV vectors.

[0086] In another embodiment, a method for generating errors in an AAV vector includes: inserting a GAA-encoding nucleic acid, or an expression cassette containing a GAA-encoding nucleic acid, as defined herein, into a packaging helper cell; and culturing the helper cells under conditions to obtain the AAV vector.

[0087] In additional embodiments, the cells are mammalian cells.

[0088] In additional embodiments, the vector-derived cells perform helper functions, such as AAV helper functions, to package the vector into a viral particle. In a particular aspect, the helper functions are performed by the Rep and/or Cap proteins for packaging the AAV vector.

[0089] In additional embodiments, cells can be stably or transiently transfected to produce a vector using polynucleotides encoding Rep and/or Cap sequences.

[0090] In additional embodiments, cells provide Rep78 and/or Rep68 proteins to obtain the vector. In the case of such cells, they can be stably or transiently transfected using polynucleotide sequences encoding Rep78 and/or Rep68 proteins.

[0091] In specific embodiments, the cells to obtain the vector are human embryonic kidney cells. In a specific aspect, the cells to produce the vector are HEK-293 cells.

Brief description of drawings

[0092] Figure 1 shows the design of pAAV-ApoE/hAAT.fixUTR.GAA13.BGH (SEQ ID NO: 16).

[0093] Figure 2 shows the design of pAAV-ApoE/hAAT.GAA13.BGH (SEQ ID NO: 17).

[0094] Figure 3 shows the design of pAAV-ApoE/hAAT.GAA7.BGH (SEQ ID NO: 18).

[0095] Figure 4 shows the design of pAAV-ApoE/hAAT.GAA8.BGH (SEQ ID NO: 19).

[0096] Figure 5 shows the design of pAAV-ApoE/hAAT.GAA2.wtBGH (SEQ ID NO: 20).

[0097] Figure 6 shows the design of pAAV-ApoE/hAAT.GAA5.wtBGH (SEQ ID NO: 21).

[0098] Figure 7 shows the design of pAAV-ApoE/hAAT.GAA7.wtBGH (SEQ ID NO: 22).

[0099] Figure 8 shows the design of pAAV-ApoE/hAAT.GAA8.wtBGH (SEQ ID NO: 23).

[0100] Figure 9 shows the design of pAAV-ApoE/hAAT.GAA13.wtBGH. (SEQ ID NO: 24)

[0101] Figure 10 shows the assessment of GAA activity levels in Huh7 cell supernatants by GAA activity assay.

[0102] Figure 11 shows the assessment of GAA activity levels in mouse plasma by GAA activity assay.

[0103] Figure 12 shows the assessment of GAA levels in mouse plasma by GAA Western blotting.

[0104] Figure 13 shows the assessment of GAA activity levels in mouse plasma by GAA activity assay.

[0105] Figure 14 shows the assessment of GAA levels in mouse plasma by GAA Western blotting.

[0106] Figure 15 shows the assessment of GAA activity levels in mouse plasma by GAA activity assay.

[0107] Figure 16 shows the assessment of GAA activity levels in rat and mouse plasma by GAA activity assay.

[0108] Figure 17 shows the assessment of GAA activity levels in plasma of non-human primates (male rhesus monkeys) by GAA activity assay.

[0109] Figure 18 shows the assessment of GAA activity levels in plasma of a non-human primate (green monkey) by GAA activity assay. The x-axis represents days after injection.

[0110] Figure 19 shows an assessment of plasma GAA levels in a non-human primate (green monkey) by Western blotting, with a graphical representation of some of the data.

[0111] Figure 20 shows an assessment of GAA activity in supernatants of Huh7 cells transduced with SPK-AAV-01, SPK-AAV-02 or a negative control (rAAV with a non-GAA transgene packaged in an AAV vector SEQ ID NO: 30- 32).

[0112] Figure 21 shows an assessment of GAA activity in the plasma of rhesus macaques after a single intravenous administration of SPK-AAV-02 at doses of 2×10 ¹² , 6×10 ¹² , 2×10 ¹³ VG/kg.

Detailed description

[0113] The invention relates to modified GAA-encoding nucleic acids, expression cassettes containing modified GAA-encoding nucleic acids, AAV vector genomes containing modified GAA-encoding nucleic acids, and recombinant AAV vectors and particles containing modified GAA-encoding nucleic acids . Modified GAA-encoding nucleic acids, expression cassettes containing modified GAA-encoding nucleic acids, AAV vector genomes containing modified GAA-encoding nucleic acids, and recombinant AAV vectors and particles of the invention can be used to treat Pompe disease as well as other glycogen diseases ( GSD).

[0114] As used herein, the term “modify” and its grammatical variants mean that a nucleic acid or protein deviates from a reference or parental sequence. The modified GAA-encoding nucleic acid is altered from a reference (eg, wild-type) or parent nucleic acid. Thus, modified nucleic acids may have substantially the same, greater or lesser activity or function with respect to the reference or parent nucleic acid, but at least retain partial activity, function and or sequence identity with respect to the reference or parent nucleic acid. The modified nucleic acid can be genetically modified to encode a modified GAA or a variant of GAA.

[0115] The term "modified GAA-encoding nucleic acid" means that the GAA nucleic acid has a change compared to the parent unmodified GAA-encoding nucleic acid. A specific example of a modification is a nucleotide substitution. The term "modification" in the present description may not be mentioned in every case where reference is made to a nucleic acid encoding a GAA.

[0116] In specific embodiments, in the case of a modified GAA-encoding nucleic acid, the GAA protein retains at least a portion of the function or activity of the wild-type GAA protein. The function or activity of the GAA protein includes the activity of acid alpha-glucosidase, a lysosomal hydrolase leading to the degradation of glycogen, maltose and isomaltose. Thus, modified GAA-encoding nucleic acids include modified forms provided that the encoded GAA retains some degree or aspect of lysosomal GAA hydrolase activity.

[0117] As discussed herein, modified GAA-encoding nucleic acids may exhibit different features or characteristics compared to the reference or parent nucleic acid. For example, modified nucleic acids include sequences with 100% identity to a reference GAA-encoding nucleic acid as defined herein, as well as sequences with less than 100% identity to a reference GAA-encoding nucleic acid.

[0118] The terms “identity”, “homology” and grammatical variants thereof mean that two or more of the substances mentioned are the same when they are “aligned” sequences. Thus, by way of example, if two nucleic acids are identical, they have the same sequence at least within the specified region or portion. Identity may extend to a specific region (or domain) of sequence.

[0119] The term "region" of identity refers to the parts of two or more specified substances that are the same. Thus, if two protein or nucleic acid sequences are identical in one or more sequence regions, they have identity in that region. The term "aligned" sequence refers to a set of protein (amino acid) or nucleic acid sequences, often containing corrections for missing or additional bases or amino acids (gaps) compared to the reference sequence.

[0120] The identity may extend over the entire length of the sequence or part of the sequence. In some embodiments, the length of the sequence having percent identity is 2, 3, 4, 5 or more contiguous amino acids or nucleic acids, for example, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 18, 19, 20, etc. adjacent nucleic acids or amino acids. In further embodiments, the length of the sequence having the identity is 21 or more contiguous amino acids or nucleic acids, for example, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, etc. adjacent amino acids or nucleic acids. In further embodiments, the length of the sequence having the identity is 41 or more contiguous amino acids or nucleic acids, for example, 42, 43, 44, 45, 45, 47, 48, 49, 5, etc. adjacent amino acids or nucleic acids. In further embodiments, the length of the sequence having the identity is 50 or more contiguous amino acids or nucleic acids, for example, 50-55, 55-60, 60-65, 65-70, 70-75, 75-80, 80-85, 85-90, 90-95, 95-100, 100-150, 150-200, 200-250, 250-300, 300-500, 500-1000, etc. adjacent amino acids or nucleic acids.

[0121] As described herein, the modified GAA-encoding nucleic acids may be different from or exhibit 100% identity or less than 100% identity to the reference GAA-encoding nucleic acid.

[0122] In certain embodiments, the GAA-encoding nucleic acids of the invention may be greater than 86% identical to any of the nucleic acids set forth in SEQ ID NOs: 1-5.

[0123] In certain embodiments, the GAA-encoding nucleic acids of the invention may be more than 87% identical to any of the nucleic acids set forth in SEQ ID NOs: 6-14.

[0124] In certain embodiments, the GAA-encoding nucleic acids of the invention may be more than 91% identical to any of the nucleic acids set forth in SEQ ID NOs: 16-24.

[0125] Such modified GAA-encoding nucleic acids may exhibit even greater identity, for example being more than 87% identical; more than 88% identical, more than 89% identical, more than 90% identical, more than 91% identical, more than 92% identical, more than 93% identical, more than 94% identical, more more than 95% identical, more than 96% identical, more than 97% identical, more than 98% identical, more than 99% identical, or 100% identical with respect to any of SEQ ID NO: 1-24.

[0126] The degree of identity (homology) or "percentage identity" between two sequences can be determined using a computer program and/or a mathematical algorithm. For purposes of the present invention, comparisons of nucleic acid sequences are made using the GCG Wisconsin Package version 9.1 software package available from the Genetics Computer Group in Madison, Wisconsin. For convenience, the comparison of sequence identity within the scope of the present invention uses the default parameters (open gap penalty = 12, gap extension penalty = 4) defined for this program. Alternatively, the Blastn 2.0 program provided by the National Center for Biotechnology Information (available online at ncbi.nlm.nih.gov/blast/; Altschul et al .) can be used to determine the level of identity and similarity between nucleic acid sequences and amino acid sequences. , 1990, J Mol Biol 215:403-410) using gap alignment using default parameters. In the case of polypeptide sequence comparisons, the BLASTP algorithm is typically used in combination with a weight matrix such as PAM100, PAM 250, BLOSUM 62 or BLOSUM 50. For quantitative analysis of the degree of identity, FASTA sequence comparison programs are also used (for example, FASTA2 and FASTA3 ) and SSEARCH (Pearson et al ., Proc. Natl. Acad. Sci. USA 85:2444 (1988); Pearson, Methods Mol Biol. 132:185 (2000) and Smith et al ., J. Mol. Biol. 147 :195 (1981)). Programs have also been developed to quantify the structural similarity of proteins using topological mapping based on Delaunay triangulation (Bostick et al ., Biochem Biophys Res Commun. 304:320 (2003)).

[0127] Modified GAA-encoding nucleic acids exhibiting different features or characteristics compared to the reference or parent nucleic acid include nucleotide substitutions. For example, modified GAA-encoding nucleic acids include nucleic acids with a reduced number of CpG dinucleotides compared to the reference GAA-encoding nucleic acid.

[0128] In some embodiments, the GAA-encoding nucleic acid contains less than 127 CpG dinucleotides.

[0129] In some embodiments, the GAA-encoding nucleic acid contains less than 126 CpG dinucleotides.

[0130] In some embodiments, the GAA-encoding nucleic acid contains less than 125 CpG dinucleotides.

[0131] In some embodiments, the GAA-encoding nucleic acid contains less than 124 CpG dinucleotides.

[0132] In some embodiments, the GAA-encoding nucleic acid contains less than 123 CpG dinucleotides.

[0133] In some embodiments, the GAA-encoding nucleic acid contains less than 122 CpG dinucleotides.

[0134] In some embodiments, the GAA-encoding nucleic acid contains less than 121 CpG dinucleotides.

[0135] In some embodiments, the GAA-encoding nucleic acid contains less than 120 CpG dinucleotides.

[0136] In some embodiments, the GAA-encoding nucleic acid contains less than 119 CpG dinucleotides.

[0137] In some embodiments, the GAA-encoding nucleic acid contains less than 118 CpG dinucleotides.

[0138] In some embodiments, the GAA-encoding nucleic acid contains less than 117 CpG dinucleotides.

[0139] In some embodiments, the GAA-encoding nucleic acid contains less than 116 CpG dinucleotides.

[0140] In some embodiments, the GAA-encoding nucleic acid contains less than 115 CpG dinucleotides.

[0141] In some embodiments, the GAA-encoding nucleic acid contains less than 114 CpG dinucleotides.

[0142] In some embodiments, the GAA-encoding nucleic acid contains less than 113 CpG dinucleotides.

[0143] In some embodiments, the GAA-encoding nucleic acid contains less than 112 CpG dinucleotides.

[0144] In some embodiments, the GAA-encoding nucleic acid contains less than 111 CpG dinucleotides.

[0145] In some embodiments, the GAA-encoding nucleic acid contains less than 110 CpG dinucleotides.

[0146] In some embodiments, the GAA-encoding nucleic acid contains less than 109 CpG dinucleotides.

[0147] In some embodiments, the GAA-encoding nucleic acid contains less than 108 CpG dinucleotides.

[0148] In some embodiments, the GAA-encoding nucleic acid contains less than 107 CpG dinucleotides.

[0149] In some embodiments, the GAA-encoding nucleic acid contains less than 106 CpG dinucleotides.

[0150] In some embodiments, the GAA-encoding nucleic acid contains less than 105 CpG dinucleotides.

[0151] In some embodiments, the GAA-encoding nucleic acid contains less than 104 CpG dinucleotides.

[0152] In some embodiments, the GAA-encoding nucleic acid contains less than 103 CpG dinucleotides.

[0153] In some embodiments, the GAA-encoding nucleic acid contains less than 102 CpG dinucleotides.

[0154] In some embodiments, the GAA-encoding nucleic acid contains less than 101 CpG dinucleotides.

[0155] In some embodiments, the GAA-encoding nucleic acid contains less than 100 CpG dinucleotides.

[0156] In some embodiments, the GAA-encoding nucleic acid contains less than 99 CpG dinucleotides.

[0157] In some embodiments, the GAA-encoding nucleic acid contains less than 98 CpG dinucleotides.

[0158] In some embodiments, the GAA-encoding nucleic acid contains less than 97 CpG dinucleotides.

[0159] In some embodiments, the GAA-encoding nucleic acid contains less than 96 CpG dinucleotides.

[0160] In some embodiments, the GAA-encoding nucleic acid contains less than 95 CpG dinucleotides.

[0161] In some embodiments, the GAA-encoding nucleic acid contains less than 94 CpG dinucleotides.

[0162] In some embodiments, the GAA-encoding nucleic acid contains less than 93 CpG dinucleotides.

[0163] In some embodiments, the GAA-encoding nucleic acid contains less than 92 CpG dinucleotides.

[0164] In some embodiments, the GAA-encoding nucleic acid contains less than 91 CpG dinucleotides.

[0165] In some embodiments, the GAA-encoding nucleic acid contains less than 90 CpG dinucleotides.

[0166] In some embodiments, the GAA-encoding nucleic acid contains less than 89 CpG dinucleotides.

[0167] In some embodiments, the GAA-encoding nucleic acid contains less than 88 CpG dinucleotides.

[0168] In some embodiments, the GAA-encoding nucleic acid contains less than 87 CpG dinucleotides.

[0169] In some embodiments, the GAA-encoding nucleic acid contains less than 86 CpG dinucleotides.

[0170] In some embodiments, the GAA-encoding nucleic acid contains less than 85 CpG dinucleotides.

[0171] In some embodiments, the GAA-encoding nucleic acid contains less than 84 CpG dinucleotides.

[0172] In some embodiments, the GAA-encoding nucleic acid contains less than 83 CpG dinucleotides.

[0173] In some embodiments, the GAA-encoding nucleic acid contains less than 82 CpG dinucleotides.

[0174] In some embodiments, the GAA-encoding nucleic acid contains less than 81 CpG dinucleotides.

[0175] In some embodiments, the GAA-encoding nucleic acid contains less than 80 CpG dinucleotides.

[0176] In some embodiments, the GAA-encoding nucleic acid contains less than 79 CpG dinucleotides.

[0177] In some embodiments, the GAA-encoding nucleic acid contains less than 78 CpG dinucleotides.

[0178] In some embodiments, the GAA-encoding nucleic acid contains less than 77 CpG dinucleotides.

[0179] In some embodiments, the GAA-encoding nucleic acid contains less than 76 CpG dinucleotides.

[0180] In some embodiments, the GAA-encoding nucleic acid contains less than 75 CpG dinucleotides.

[0181] In some embodiments, the GAA-encoding nucleic acid contains less than 74 CpG dinucleotides.

[0182] In some embodiments, the GAA-encoding nucleic acid contains less than 73 CpG dinucleotides.

[0183] In some embodiments, the GAA-encoding nucleic acid contains less than 72 CpG dinucleotides.

[0184] In some embodiments, the GAA-encoding nucleic acid contains less than 71 CpG dinucleotides.

[0185] In some embodiments, the GAA-encoding nucleic acid contains less than 70 CpG dinucleotides.

[0186] In some embodiments, the GAA-encoding nucleic acid contains less than 69 CpG dinucleotides.

[0187] In some embodiments, the GAA-encoding nucleic acid contains less than 68 CpG dinucleotides.

[0188] In some embodiments, the GAA-encoding nucleic acid contains less than 67 CpG dinucleotides.

[0189] In some embodiments, the GAA-encoding nucleic acid contains less than 66 CpG dinucleotides.

[0190] In some embodiments, the GAA-encoding nucleic acid contains less than 65 CpG dinucleotides.

[0191] In some embodiments, the GAA-encoding nucleic acid contains less than 64 CpG dinucleotides.

[0192] In some embodiments, the GAA-encoding nucleic acid contains less than 63 CpG dinucleotides.

[0193] In some embodiments, the GAA-encoding nucleic acid contains less than 62 CpG dinucleotides.

[0194] In some embodiments, the GAA-encoding nucleic acid contains less than 61 CpG dinucleotides.

[0195] In some embodiments, the GAA-encoding nucleic acid contains less than 60 CpG dinucleotides.

[0196] In some embodiments, the GAA-encoding nucleic acid contains less than 59 CpG dinucleotides.

[0197] In some embodiments, the GAA-encoding nucleic acid contains less than 58 CpG dinucleotides.

[0198] In some embodiments, the nucleic acid encoding the GAA contains less than 57 CpG dinucleotides.

[0199] In some embodiments, the GAA-encoding nucleic acid contains less than 56 CpG dinucleotides.

[0200] In some embodiments, the GAA-encoding nucleic acid contains less than 55 CpG dinucleotides.

[0201] In some embodiments, the GAA-encoding nucleic acid contains less than 54 CpG dinucleotides.

[0202] In some embodiments, the GAA-encoding nucleic acid contains less than 53 CpG dinucleotides.

[0203] In some embodiments, the nucleic acid encoding the GAA contains less than 52 CpG dinucleotides.

[0204] In some embodiments, the GAA-encoding nucleic acid contains less than 51 CpG dinucleotides.

[0205] In some embodiments, the GAA-encoding nucleic acid contains less than 50 CpG dinucleotides.

[0206] In some embodiments, the GAA-encoding nucleic acid contains less than 49 CpG dinucleotides.

[0207] In some embodiments, the GAA-encoding nucleic acid contains less than 48 CpG dinucleotides.

[0208] In some embodiments, the GAA-encoding nucleic acid contains less than 47 CpG dinucleotides.

[0209] In some embodiments, the GAA-encoding nucleic acid contains less than 46 CpG dinucleotides.

[0210] In some embodiments, the GAA-encoding nucleic acid contains less than 45 CpG dinucleotides.

[0211] In some embodiments, the GAA-encoding nucleic acid contains less than 44 CpG dinucleotides.

[0212] In some embodiments, the GAA-encoding nucleic acid contains less than 43 CpG dinucleotides.

[0213] In some embodiments, the GAA-encoding nucleic acid contains less than 42 CpG dinucleotides.

[0214] In some embodiments, the GAA-encoding nucleic acid contains less than 41 CpG dinucleotides.

[0215] In some embodiments, the GAA-encoding nucleic acid contains less than 40 CpG dinucleotides.

[0216] In some embodiments, the GAA-encoding nucleic acid contains less than 39 CpG dinucleotides.

[0217] In some embodiments, the GAA-encoding nucleic acid contains less than 38 CpG dinucleotides.

[0218] In some embodiments, the GAA-encoding nucleic acid contains less than 37 CpG dinucleotides.

[0219] In some embodiments, the GAA-encoding nucleic acid contains less than 36 CpG dinucleotides.

[0220] In some embodiments, the GAA-encoding nucleic acid contains less than 35 CpG dinucleotides.

[0221] In some embodiments, the GAA-encoding nucleic acid contains less than 34 CpG dinucleotides.

[0222] In some embodiments, the GAA-encoding nucleic acid contains less than 33 CpG dinucleotides.

[0223] In some embodiments, the GAA-encoding nucleic acid contains less than 32 CpG dinucleotides.

[0224] In some embodiments, the GAA-encoding nucleic acid contains less than 31 CpG dinucleotides.

[0225] In some embodiments, the nucleic acid encoding the GAA contains less than 30 CpG dinucleotides.

[0226] In some embodiments, the GAA-encoding nucleic acid contains less than 29 CpG dinucleotides.

[0227] In some embodiments, the GAA-encoding nucleic acid contains less than 28 CpG dinucleotides.

[0228] In some embodiments, the GAA-encoding nucleic acid contains less than 27 CpG dinucleotides.

[0229] In some embodiments, the GAA-encoding nucleic acid contains less than 26 CpG dinucleotides.

[0230] In some embodiments, the GAA-encoding nucleic acid contains less than 25 CpG dinucleotides.

[0231] In some embodiments, the GAA-encoding nucleic acid contains less than 24 CpG dinucleotides.

[0232] In some embodiments, the GAA-encoding nucleic acid contains less than 23 CpG dinucleotides.

[0233] In some embodiments, the GAA-encoding nucleic acid contains less than 22 CpG dinucleotides.

[0234] In some embodiments, the GAA-encoding nucleic acid contains less than 21 CpG dinucleotides.

[0235] In some embodiments, the GAA-encoding nucleic acid contains less than 20 CpG dinucleotides.

[0236] In some embodiments, the GAA-encoding nucleic acid contains less than 19 CpG dinucleotides.

[0237] In some embodiments, the GAA-encoding nucleic acid contains less than 18 CpG dinucleotides.

[0238] In some embodiments, the GAA-encoding nucleic acid contains less than 17 CpG dinucleotides.

[0239] In some embodiments, the GAA-encoding nucleic acid contains less than 16 CpG dinucleotides.

[0240] In some embodiments, the GAA-encoding nucleic acid contains less than 15 CpG dinucleotides.

[0241] In some embodiments, the GAA-encoding nucleic acid contains less than 14 CpG dinucleotides.

[0242] In some embodiments, the GAA-encoding nucleic acid contains less than 13 CpG dinucleotides.

[0243] In some embodiments, the GAA-encoding nucleic acid contains less than 12 CpG dinucleotides.

[0244] In some embodiments, the nucleic acid encoding the GAA contains less than 11 CpG dinucleotides.

[0245] In some embodiments, the GAA-encoding nucleic acid contains less than 10 CpG dinucleotides.

[0246] In some embodiments, the nucleic acid encoding the GAA contains less than 9 CpG dinucleotides.

[0247] In some embodiments, the nucleic acid encoding the GAA contains less than 8 CpG dinucleotides.

[0248] In some embodiments, the GAA-encoding nucleic acid contains less than 7 CpG dinucleotides.

[0249] In some embodiments, the GAA-encoding nucleic acid contains less than 6 CpG dinucleotides.

[0250] In some embodiments, the GAA-encoding nucleic acid contains less than 5 CpG dinucleotides.

[0251] In some embodiments, the GAA-encoding nucleic acid contains less than 4 CpG dinucleotides.

[0252] In some embodiments, the GAA-encoding nucleic acid contains less than 3 CpG dinucleotides.

[0253] In some embodiments, the GAA-encoding nucleic acid contains less than 2 CpG dinucleotides.

[0254] In some embodiments, the nucleic acid encoding the GAA contains less than 1 CpG dinucleotide.

[0255] The term “vector” refers to a small nucleic acid molecule carrier, plasmid, virus (eg, AAV vector) or other carrier that can be manipulated by insertion or integration of a nucleic acid. Such vectors can be used for genetic manipulation (ie, "cloning vectors"), for insertion/transfer of polynucleotides into cells, and for transcription or translation of the inserted polynucleotide in cells. An "expression vector" is a special vector containing a gene or nucleic acid sequence with regulatory regions required for expression in a host cell.

[0256] The vector nucleic acid sequence typically comprises at least an origin of replication for propagation in the cell and optionally additional elements such as a heterologous polynucleotide sequence, an expression control element (e.g., promoter, enhancer), an intron, an inverted terminal repeat (ITR), selectable marker (eg, antibiotic resistance), polyadenylation signal.

[0257] A viral vector is derived from or is based on one or more nucleic acid elements comprising a viral genome. Specific viral vectors include lentiviral vectors and adeno-associated virus (AAV) vectors.

[0258] The term "recombinant" as a definition of a vector, such as a recombinant AAV vector (rAAV), as well as a definition of sequences, such as recombinant polynucleotides and polypeptides, means that the compositions have been acted upon (i.e., engineered) in a manner that As a rule, this does not occur in nature. A specific example of a recombinant AAV vector would be a vector in which a click reaction nucleic acid sequence that is not normally present in the wild-type AAV genome is inserted into the AAV genome. Although the term “recombinant” is not always used herein to refer to AAV vectors as well as sequences such as polynucleotides, recombinant forms including polynucleotides are specifically included herein, notwithstanding any such omission.

[0259] A "recombinant AAV vector" or "rAAV" is prepared from the wild-type AAV genome using molecular techniques to remove the wild-type genome from the AAV genome and replace it with a non-native nucleic acid sequence, referred to as a heterologous nucleic acid. Typically, in the case of AAV, one or both inverted terminal repeat (ITR) sequences of the AAV genome are retained in the AAV vector. rAAV is different from the AAV genome because the entire AAV genome or a portion thereof is replaced with a non-native sequence relative to the AAV genomic nucleic acid. Thus, the insertion of a non-native sequence defines the AAV vector as a “recombinant” vector, which can be referred to as a “rAAV vector”.

[0260] The rAAV sequence can be packaged, in which case it is referred to herein as a "particle", for subsequent infection (transduction) of a cell ex vivo , in vitro or in vivo . If the recombinant AAV vector sequence is encapsided or packaged into an AAV particle, the particle may also be referred to as an “rAAV vector” or “rAAV particle.” Such rAAV particles include proteins that enclose the capsid or package the vector genome and, in the case of AAV, are referred to as capsid proteins.

[0261] The term vector "genome" refers to the portion of the recombinant plasmid sequence that is ultimately packaged or encapsided to form a viral particle (eg, rAAV). In cases where recombinant plasmids are used for the design or production of recombinant vectors, the vector genome does not include a portion of the “plasmid” that does not correspond to the vector genome sequence of the recombinant plasmid. This portion of the recombinant plasmid, which is not the vector genome, can be referred to as the "plasmid backbone", which is important for cloning and amplification of the plasmid, a process necessary for the propagation and production of the recombinant virus, but is not itself packaged or capsided into viral particles ( e.g. AAV). Thus, the term vector "genome" refers to the nucleic acid packaged or encapsided by a virus (eg, AAV).

[0262] Host cells for producing recombinant AAV particles include, but are not limited to, microorganisms, yeast cells, insect cells, and mammalian cells that can be used or are used as recipients of heterologous rAAV vectors. Cells from the stable human cell line HEK293 (available, for example, from the American Type Culture Collection under ATCC accession number CRL1573) can be used. In some embodiments, a modified human embryonic kidney cell line (eg, HEK293) transformed with adenovirus type-5 DNA fragments and expressing adenoviral E1a and E1b genes is used to produce recombinant AAV particles. The modified HEK293 cell line is easy to transfect and provides a particularly convenient platform for the production of rAAV particles. Other host cell lines suitable for recombinant AAV production are described in international application PCT/2017/024951.

[0263] In some embodiments, AAV helper functions are introduced into a host cell by transfecting the host cell with an AAV helper construct prior to or simultaneously with transfection of an AAV expression vector. A host cell having AAV helper functions may be referred to as a "helper cell" or a "packaging helper cell". Thus, AAV helper constructs are sometimes used to provide at least transient expression of the AAV rep and/or cap genes to supplement the missing AAV functions required for productive AAV transduction. AAV helper constructs often lack AAV ITRs and cannot replicate or package themselves. These constructs may be in the form of a plasmid, phage, transposon, cosmid, virus or virion. A number of AAV helper constructs have been described, such as the widely used plasmids pAAV/Ad and pIM29+45, encoding the expression products of Rep and Cap. A number of other vectors encoding expression products of Rep and/or Cap are known.

[0264] Methods are known in the art for producing recombinant AAV particles that can be used to transduce mammalian cells. For example, recombinant AAV particles can be produced as described in US Pat. No. 9,408,904 and International Application Nos. PCT/US2017/025396 and PCT/US2016/064414.

[0265] The terms “nucleic acid” and “polynucleotide” are used interchangeably herein to refer to all forms of nucleic acid, oligonucleotides, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Nucleic acids include genomic DNA, cDNA and antisense DNA and spliced or unspliced mRNA, rRNA, tRNA and inhibitory DNA or RNA (RNAi, e.g. small or short hairpin (sh)RNA, microRNA (miRNA), small or short interfering (mi) RNA, trans-spliced RNA or antisense RNA). Nucleic acids include natural, synthetic and intentionally modified or altered polynucleotides (eg, variant nucleic acid). Nucleic acids such as cDNA, genomic DNA, RNA and fragments thereof may be single or double stranded.

[0266] Polynucleotides can be single-stranded, double-stranded or triplex, linear or circular, and can be of any length. In the polynucleotides discussed, the sequence or structure of a particular polynucleotide can be described in accordance with the sequence representation in the 5'-3' direction.

[0267] The term “transgene” is used herein to refer to a heterologous nucleic acid intended to be inserted or incorporated into a cell or organism. Transgenes include any heterologous nucleic acid, such as a modified nucleic acid encoding GAA.

[0268] The term “transduce” and its grammatical variations refer to the insertion of a molecule, such as an rAAV vector, into a cell or host organism. The heterologous nucleic acid/transgene may or may not integrate into the genomic nucleic acid of the recipient cell. The inserted heterologous nucleic acid may also exist in the recipient cell or host organism extrachromosomally or only transiently.

[0269] A "transduced cell" is a cell into which a transgene has been inserted. Thus, the term "transduced" cell (eg, in a mammal, such as a cell, or tissue, or organ cell) means a genetic change in the cell following the insertion, for example, of a nucleic acid (eg, a transgene) into the cell. Thus, a "transduced" cell is a cell into which, or into its progeny, an exogenous nucleic acid has been inserted. Cells can be grown and the integrated protein can be expressed. When used in gene therapy and gene therapy methods, the transduced cell may be present in an individual.

[0270] The term “expression control element” refers to nucleic acid sequences that influence the expression of an operably linked nucleic acid. Expression control elements as defined herein include promoters and enhancers. Vector sequences, including AAV vectors, may include one or more “expression control elements.” Typically, such elements are included to facilitate the correct transcription of a heterologous polynucleotide and, if necessary, its translation (e.g., a promoter, an enhancer, a splicing signal for introns, maintaining the correct reading frame of the gene for translation of in-frame mRNA and stop codons, etc.). d.). Such elements typically act in cis and are referred to as “cis-acting” elements, but they can also act in trans.

[0271] Expression control can be influenced at the level of transcription, translation, splicing, mRNA stability, etc. Typically, an expression control element that modulates transcription is located near the 5' end (ie, "upstream") of the transcribed nucleic acid. Expression control elements may also be located at the 3' end (ie, "downstream") of the transcribed sequence or within the transcript (eg, in an intron). Expression control elements may be contiguous or distant from the transcribed sequence (eg, 1-10, 10-25, 25-50, 50-100, 100 to 500 or more nucleotides from the polynucleotide), even at a significant distance. However, due to the length limitations of AAV vectors, expression control elements will typically be located within 1 to 1000 nucleotides of the transcription start site of a heterologous nucleic acid.

[0272] Functionally, the expression of an operably linked nucleic acid is at least partially controlled by an element (eg, a promoter) such that the element modulates transcription of the nucleic acid and, if necessary, translation of the transcript. A specific example of an expression control element is a promoter, typically located in the 5' direction of the transcribed nucleic acid sequence. A promoter generally increases the amount of product expressed from the operably linked nucleic acid relative to the amount expressed in the absence of a promoter.

[0273] As used herein, the term “enhancer” may refer to a sequence adjacent to a heterologous nucleic acid. Enhancer elements are typically found upstream of a promoter element, but also function and can be found downstream or within the sequence. Thus, the enhancer element may be 10-50 base pairs, 50-100 base pairs, 100-200 base pairs, or 200-300 base pairs or more base pairs upstream or downstream of the heterologous nucleic acid sequence. Enhancer elements typically increase the expression of an operably linked nucleic acid above the expression provided by the promoter element.

[0274] The expression construct may contain regulatory elements that serve to regulate expression in a particular cell or tissue type. Expression control elements (eg, promoters) include elements active in a particular tissue or cell type, referred to herein as "tissue-specific expression control elements/promoters." Tissue-specific expression control elements are typically active in a specific cell or tissue (eg, liver). Expression control elements are typically active in specific cells, tissues, or organs because they are recognized by transcriptional activator proteins or other transcriptional regulators that are unique to a particular cell type, tissue, or organ. Such regulatory elements are known to those skilled in the art (see, for example, Sambrook et al . (1989) and Ausubel et al . (1992)).

[0275] The inclusion of tissue-specific regulatory elements in expression constructs provides at least partial tissue tropism for the expression of a heterologous nucleic acid encoding a protein or inhibitory RNA. Examples of promoters that are active in the liver include, but are not limited to, the transthyretin (TTR) gene promoter; human 1-antitrypsin alpha (hAAT) promoter; albumin promoter, Miyatake, et al ., J. Virol. , 71:5124-32 (1997); hepatitis B virus core promoter, Sandig, et al ., Gene Ther . 3:1002-9 (1996); alpha-fetoprotein (AFP) promoter, Arbuthnot, et al ., Hum. Gene. Ther ., 7:1503-14 (1996). An example of an enhancer active in the liver is apolipoprotein E (apoE) HCR-1 and HCR-2 (Allan et al ., J. Biol. Chem. , 272:29113-19 (1997)).

[0276] Expression control elements also include ubiquitous or promiscuous promoters/enhancers capable of regulating polynucleotide expression in a variety of different cell types. Such elements include, but are not limited to, cytomegalovirus (CMV) immediate early promoter/enhancer sequences, Rous sarcoma virus (RSV) promoter/enhancer sequences, and other viral promoters/enhancers active in various mammalian cell types, or synthetic elements not found in nature. (cm., for example Boshart et al., Cell, 41:521-530 (1985)), SV40 promoter, dihydrofolate reductase promoter, cytoplasmic β-actin promoter and phosphoglycerol kinase (PGK) promoter.

[0277] Expression control elements can also provide expression in a controlled manner, i.e. a signal or stimuli increases or decreases the expression of an operably linked heterologous polynucleotide. A regulated element that increases the expression of an operably linked polynucleotide in response to a signal or stimuli is also referred to as an "inducible element" (ie, signal inducible). Specific non-limiting examples include a hormone (eg, steroid)-inducible promoter. Typically, the degree of enhancement or reduction imparted by such elements is proportional to the magnitude of the signal or stimuli; the greater the magnitude of the signal or stimuli, the greater the increase or decrease in expression. Specific non-limiting examples include the zinc-inducible ovine metallothionein (MT) promoter; steroid-inducible mouse mammary cancer virus (MMTV) promoter; T7 polymerase promoter system (WO 98/10088); tetracycline-repressible system (Gossen, et al., Proc. Natl. Acad. Sci. USA, 89:5547-5551 (1992)); tetracycline-inducible system (Gossen, et al., Science. 268:1766-1769 (1995); also see Harvey, et al., Curr. Opin. Chem. Biol. 2:512-518 (1998)); RU486-inducible system (Wang, et al., Nat. Biotech. 15:239-243 (1997) and Wang, et al., Gene Ther. 4:432-441 (1997)]; and the rapamycin-inducible system (Magari, et al., J. Clin. Invest. 100:2865-2872 (1997); Rivera, et al., Nat. Medicine. 2:1028-1032 (1996)). Other regulated controls that can be used in this context are those regulated by a specific physiological state, for example, temperature, acute phase, developmental stage.

[0278] Expression control elements also include native elements for the heterologous polynucleotide. A native control element (eg, a promoter) can be used when it is desired that expression of a heterologous polynucleotide mimic native expression. A native element can be used when the expression of a heterologous polynucleotide is subject to regulation depending on time or developmental stage, or in a tissue-specific manner, or in response to specific transcriptional stimuli. Other native expression control elements such as introns, polyadenylation sites, or Kozak consensus sequences can also be used.

[0279] The term "operably linked" means that the regulatory sequences necessary for the expression of the nucleic acid sequence are located at suitable positions relative to the sequence so as to effect expression of the nucleic acid sequence. The same definition is sometimes used with respect to the arrangement of nucleic acid sequences and transcriptional control elements (eg, promoters, enhancers, and termination elements) in an expression vector, such as an rAAV vector.

[0280] In an example of an expression control element operably linked to a nucleic acid, the relationship is such that the control element modulates expression of the nucleic acid. More specifically, for example, the term “two operably linked DNA sequences” means that the two DNAs are arranged (cis or trans) in such a relationship that at least one of the DNA sequences can cause a physiological effect with respect to the other sequence.

[0281] Thus, additional elements for vectors include, but are not limited to, an expression control element (e.g., a promoter/enhancer), a transcription termination signal or stop codon, 5' or 3' untranslated regions (e.g., polyadenylation sequences (poly-A)) flanking sequence, such as one or more copies of the AAV ITR sequence, or an intron.

[0282] Additional elements include, for example, filler or spacer polynucleotide sequences, for example, to improve packaging and reduce the presence of contaminating nucleic acid. AAV vectors typically accept DNA inserts having a range of sizes, typically from about 4 kb. to approximately 5.2 kb. or a little more. Thus, for shorter sequences, the inclusion of a spacer or filler to adjust the length to approximately the normal or normal size of the viral genomic sequence suitable for packaging the AAV vector into a viral particle. In various embodiments, the filler/spacer nucleic acid sequence is a non-translated (non-protein coding) segment of the nucleic acid. In the case of a nucleic acid sequence less than 4.7 kb. the filler or spacer polynucleotide sequence has a length that, when combined (eg, inserted into a vector) with the sequence, has a total length of approximately 3.0-5.5 kb, or approximately 4.0-5.0 kb .b., or approximately 4.3-4.8 kb.

[0283] The term “isolated,” when used to define a composition, means that the compositions are man-made or separated wholly or at least partially from their natural in vivo environment. Typically, isolated compositions are substantially free of one or more materials with which they are typically associated in nature, eg, one or more proteins, nucleic acids, lipids, carbohydrates, cell membranes.

[0284] The term “isolated” does not exclude human-derived combinations, such as an rAAV sequence or an rAAV particle packaging or capsiding an AAV vector genome, and a pharmaceutical composition. The term “isolated” also does not exclude alternative physical forms of the composition, such as hybrids/chimeras, multimers/oligomers, modifications (eg, phosphorylation, glycosylation, lipidation), or derivatized forms or forms expressed in human host cells.

[0285] The term “substantially pure” refers to a preparation containing at least 50-60% by weight of the compound of interest (eg, nucleic acid, oligonucleotide, protein, etc.). The formulation may contain at least 75% by weight, or at least 85% by weight, or approximately 90-99% by weight, of the compound of interest. Purity is measured by methods appropriate to the compound of interest (eg, chromatographic methods, agarose or polyacrylamide gel electrophoresis, HPLC analysis, etc.).

[0286] The phrase "consisting essentially of" with respect to a particular nucleotide sequence or amino acid sequence means a sequence having the properties of the specified SEQ ID NO. For example, when used in relation to an amino acid sequence, the phrase includes the sequence itself and molecular modifications that will not affect the basic and novel characteristics of the sequence.

[0287] Nucleic acid expression vectors (eg, AAV vector genomes), plasmids, including modified GAA-encoding nucleic acids of the invention can be produced by recombinant DNA methods. The availability of nucleotide sequence information makes it possible to obtain isolated nucleic acid molecules according to the invention by various methods. Nucleic acids encoding GAAs can be produced using a variety of standard cloning techniques, recombinant DNA, cellular expression or in vitro translation, and chemical synthesis. The purity of polynucleotides can be determined by sequencing, gel electrophoresis, or the like. For example, nucleic acids can be isolated using hybridization techniques or computer database screening. Such methods include, but are not limited to: (1) hybridizing genomic DNA or cDNA libraries with probes to detect homologous nucleotide sequences; (2) screening antibodies to detect polypeptides having common structural features, for example, using an expression library; (3) polymerase chain reaction (PCR) using genomic DNA or cDNA using primers capable of annealing to the nucleic acid sequence of interest; (4) computer searches of sequence databases for related sequences; and (5) differential screening of a subtracted nucleic acid library.

[0288] Nucleic acids can be maintained as DNA in any convenient cloning vector. In one embodiment, the clones are maintained in a plasmid cloning/expression vector, such as pBluescript (Stratagene, La Jolla, Calif.), propagated in a suitable E. coli host cell. Alternatively, the nucleic acids can be maintained in a vector suitable for expression in mammalian cells, such as an AAV vector. In cases where post-translational modification affects protein function, the nucleic acid molecule can be expressed in mammalian cells.

[0289] Within the scope of the invention, rAAV vectors may optionally contain regulatory elements necessary for the expression of a heterologous nucleic acid in a cell positioned to allow expression of the encoded protein in the host cell. Such regulatory elements required for expression include, but are not limited to, promoter sequences, enhancer sequences, and transcription initiation sequences, as defined herein and known to those skilled in the art.

[0290] The methods and uses of the invention involve delivery (transduction) of a nucleic acid (transgene) into host cells, including dividing and/or non-dividing cells. The nucleic acids, rAAV vector, methods, uses and pharmaceutical compositions of the invention can further be used in a method of delivering, administering or providing a sequence encoded by a heterologous nucleic acid to an individual in need thereof as a method of treatment. Thus, the nucleic acid is transcribed and the protein is produced in vivo in the body of the individual. An individual may benefit from or need protein because... the individual has a protein deficiency, or because the production of the protein in the body of an individual may have some therapeutic effect in the form of treatment or otherwise.

[0291] The present invention can be used in animals, including humans, and in veterinary medicine. Thus, suitable individuals include mammals, such as humans, as well as non-human mammals. The term "individual" refers to an animal, typically a mammal, such as humans, non-human primates (apes, gibbons, gorillas, chimpanzees, orangutans, macaques), domestic animals (dogs and cats), farm animals (poultry, such as chickens and ducks, horses, cows, goats, sheep, pigs) and experimental animals (mice, rat, rabbit, guinea pig). Humans include fetus, newborn, infant, adolescent and adult individuals. Individuals include animal models of disease, for example, mouse and other animal models of protein/enzyme deficiencies such as Pompe disease and glycogen storage disorders (GSD), and other models known to those skilled in the art.

[0292] Individuals suitable for treatment according to the invention include individuals who produce, or are at risk of producing, insufficient amounts of GAA, or who produce abnormal, partially functional, or nonfunctional GAA. Individuals can be tested for GAA activity to determine whether such individuals are suitable for treatment with the method of the invention. Individuals suitable for treatment according to the invention also include individuals who will benefit from GAA. Such individuals who may benefit from GAA include those with glycogen storage disease (GSD). Treated individuals may be monitored after treatment periodically, for example, every 1-4 weeks, 1-6 weeks, 6-12 weeks, or 1, 2, 3, 4, 5 or more years.

[0293] Individuals can be tested for an immune response, such as anti-AAV antibodies. Thus, candidate individuals can be screened before treatment with the method of the invention. Individuals can also be tested for post-AAV antibodies after treatment and optionally monitored for a period of time after treatment. Individuals who have pre-existing or developing anti-AAV antibodies can be treated with an immunosuppressive agent or other treatment regimen as specified herein.

[0294] Individuals suitable for treatment according to the invention also include individuals who produce or are at risk of producing anti-AAV antibodies. rAAV vectors can be introduced or delivered to such individuals in several ways. For example, an empty AAV capsid (ie, an AAV lacking a modified GAA-encoding nucleic acid) can be delivered to bind to anti-AAV antibodies in an individual, thereby allowing an rAAV vector containing a heterologous nucleic acid to transduce cells of the individual.

[0295] The modified nucleic acids, rAAV expression cassettes and vectors of the invention can be used to treat GAA deficiency. Thus, in various embodiments, the modified GAA-encoding nucleic acids, expression cassettes containing the modified GAA-encoding nucleic acids, and rAAV vectors of the invention can be used as therapeutic and/or prophylactic agents.

[0296] In specific embodiments, the modified GAA-encoding nucleic acids, expression cassettes containing the modified GAA-encoding nucleic acids, and rAAV vectors of the invention can be used to treat Pompe disease as well as other glycogen diseases. Administration of modified GAA-encoding nucleic acids, expression cassettes containing modified GAA-encoding nucleic acids, and rAAV vectors of the invention to a patient with Pompe disease or other glycogen diseases results in the expression of GAA protein serving to suppress, inhibit, or reduce glycogen accumulation, preventing glycogen accumulation or glycogen degradation, which in turn may reduce or ameliorate one or more adverse effects or symptoms of Pompe disease.

[0297] Individuals, animals, or patients administered modified GAA-encoding nucleic acids, expression cassettes containing modified GAA-encoding nucleic acids, and rAAV vectors of the invention can be evaluated using a variety of tests, assays, and functional assessments to demonstrate, measure, and /or evaluating the effectiveness of the modified GAA-encoding nucleic acids, expression cassettes containing modified GAA-encoding nucleic acids, and rAAV vectors of the invention as therapeutic and/or prophylactic agents. Such tests and assays include, but are not limited to, measuring GAA activity (eg, using standard GAA activity assays) and/or amount of GAA (eg, using Western blotting using an anti-GAA antibody) in a biological sample such as blood or plasma; measuring glycogen content in tissues such as muscle tissue samples; histological evaluation of muscles (such as muscle tissue from the triceps brachii, quadriceps femoris, diaphragm and heart), spinal cord (including examination and enumeration of ChAT-positive motor neurons in the anterior horn of the cervical, thoracic and lumbar spinal cord segments and assessment of astroglial response and activation of microglia); assessment of respiratory function during quiet breathing; forelimb wire hanging test; compression force measurement; rotarod test to determine motor coordination; chest x-ray, which can detect cardiomegaly; electrocardiography (ECG) to study heart function; electromyography (EMG), which can be used to determine myopathy; measurement of GAA activity in skin fibroblasts; GAA activity assays in dried blood samples; blood/serum tests for serum creatine kinase, which, when elevated, is a nonspecific marker of Pompe disease; blood/serum tests for aminotransferase, alanine aminotransferase, or lactate dehydrogenase, which may be elevated, indicating release from muscle in Pompe disease; urine glucose tetrasaccharide assay (a sensitive, nonspecific marker of Pompe disease; analysis of peak and steady-state levels of vector-derived GAA enzyme assessed by total GAA protein and plasma activity; pulmonary function testing; muscle function testing; muscle biopsy and staining for glycogen in the vacuoles of the liver; testing for immune responses to the AAV capsid; testing for immune responses to the GAA transgene protein; testing of the forced vital capacity of the lungs; using an AAV vector of the GAA enzyme (assessed by total GAA protein and activity measured in plasma; Walking, Stair Climbing, Gowers, and Chair Rising (GSGC) test); muscle strength testing using the Medical Research Council (MRC) grading scale based on the Rasch model; patient-reported assessment of functioning/socialization; Quantitative measurements of sleep and sleep breathing through polysomnography (PSG); patient-reported measures of fatigue, daytime sleepiness, and sleep quality; Walton and Gardner-Medwin (WGM) scale; tests of respiratory function, including, as non-limiting examples, nasal inspiratory pressure (SNIP) and maximum inspiratory and expiratory pressure (MIP and MEP, respectively); liver biomarkers; β-hexosaminidase (βHexo) assay; health measures including, but not limited to, the 26-item Short Form Health Survey (SF-36); Pompe disease activity score based on the Rasch model (R-PAct); Fatigue Severity Scale (FSS); and Patient Reported Outcomes Measurement Information System (PROMIS) item banks.

[0298] In addition, modified GAA-encoding nucleic acids, expression cassettes containing modified GAA-encoding nucleic acids, and rAAV vectors of the invention can be used to treat glycogen storage disease (GSD). Glycogenoses include, for example, GSD type I (Gierke's disease), GSD type II (Pompe disease), GSD type III (Forbes disease), GSD type IV (Andersen disease, amylopectinosis), GSD type V (McArdle disease), GSD type VI (Hers disease), GSD type VII (Tarui disease) or lethal congenital cardiac glycogenosis.

[0299] As discussed herein, rAAVs can be used as gene therapy vectors because they can enter cells and insert nucleic acid/genetic material into cells. Because AAVs are not associated with disease in humans, and rAAV vectors can deliver heterologous polynucleotide sequences (eg, therapeutic proteins and agents) to human patients without causing significant pathology or disease associated with AAV.

[0300] rAAV vectors have a number of desirable features for such use, including tropism for dividing and non-dividing cells. Early clinical experience with these vectors has also shown no consistent toxicity, and immune responses are typically minimal or undetectable. AAV is known to infect a wide range of cell types in vivo via receptor-mediated endocytosis and transcytosis. These vector systems have been tested in humans, targeting many tissues such as retinal epithelium, liver, skeletal muscle, airway, brain, joints, and hematopoietic stem cells.

[0301] It may be desirable to insert an rAAV vector that can provide, for example, multiple copies of GAA and thus higher amounts of GAA protein. Improved rAAV vectors and methods for producing these vectors are described in detail in a number of references, patents and patent applications, including Wright J.F. (Hum. Gene Ther., 20:698-706, 2009).

[0302] Direct delivery of rAAV vectors or ex vivo transduction of human cells followed by infusion into the body results in the expression of heterologous nucleic acid, thereby causing a beneficial therapeutic effect of hemostasis. In the context of modified GAA-encoding nucleic acids, administration suppresses, inhibits, or reduces the amount or accumulation of glycogen, prevents glycogen accumulation, or results in glycogen degradation. This, in turn, may result in the suppression, inhibition, reduction or reduction of one or more of the adverse effects of Pompe disease, for example, stimulation or improvement of muscle tone and/or muscle strength and/or reduction of an enlarged liver.

[0303] The recombinant AAV vector, methods and uses thereof include any strain or serotype of the virus. As a non-limiting example, the recombinant AAV vector can be based on any AAV genome, such as AAV (SEQ ID NO: 30-32), LK03 (SEQ ID NO: 33), AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, Rh10, Rh74, AAV3B or AAV-2i8, for example. Such vectors may be based on the same strain or serotype (or subgroup or variant) or may differ from each other. As a non-limiting example, a recombinant AAV vector based on the genome of a particular serotype may be identical to the serotype of the capsid proteins into which the vector is packaged. In addition, the recombinant AAV vector genome may be based on the genome of an AAV serotype that is different from the serotype of the AAV capsid proteins in which the vector is packaged. For example, the AAV vector genome may be based on AAV2, while at least one of the three capsid proteins may be, for example, AAV (SEQ ID NO: 30-32), LK03 (SEQ ID NO: 33), AAV1 , AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, Rh10, Rh74, AAV3B or AAV-2i8 or a variant thereof.

[0304] In specific embodiments, adeno-associated virus (AAV) vectors include AAV (SEQ ID NO: 30-32), LK03 (SEQ ID NO: 33), AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, Rh10, Rh74, AAV3B and AAV-2i8, as well as variants thereof (e.g. capsid variants such as insertions, additions, substitutions and deletions of amino acids), for example as described in WO 2013/158879 ( International Application No. PCT/US2013/037170), WO 2015/013313 (International Application No. PCT/US2014/047670) and US 2013/0059732 (US Patent No. 9169299 describes LK01, LK02, LK03, etc.).

[0305] As used herein, the term “serotype” is a distinctive term used to designate an AAV having a capsid that is serologically distinct from other AAV serotypes. Serological signatures are determined based on the lack of cross-reactivity between antibodies against one AAV versus another AAV. Such differences in cross-reactivity are typically the result of capsid protein sequences/antigenic determinants (eg, due to differences in the VP1, VP2 and/or VP3 sequence of AAV serotypes). Although it is possible that AAV variants, including capsid variants, may not be serologically different from the reference AAV or another AAV serotype, they differ in at least one nucleotide or amino acid residue from the reference or another AAV serotype.

[0306] According to the generally accepted definition, the term "serotype" means that the virus of interest was tested against serum specific for all existing and characterized serotypes for neutralizing activity, and no antibodies were detected that neutralize the virus of interest. As more and more natural virus isolates are discovered and/or more capsid mutants are obtained, serological differences from any of the currently existing serotypes may or may not occur. Thus, in cases where a new virus (eg, AAV) is not serologically different, the new virus (eg, AAV) will be a subgroup or variant of the corresponding serotype. In many cases, serological testing for neutralizing activity has yet to be performed on mutant viruses with modifications to the capsid sequences to determine whether they belong to a different serotype according to the generally accepted definition of serotype. Thus, for brevity, convenience, and to avoid repetition, the term "serotype" broadly refers to serologically distinct viruses (e.g., AAV) as well as serologically undifferentiated viruses (e.g., AAV), which may be within a subgroup or variant the specified serotype.

[0307] As discussed herein, AAV capsid proteins may exhibit less than 100% sequence identity to the reference or parental AAV serotype, such as AAV (SEQ ID NO: 30-32), LK03 (SEQ ID NO: 33), AAV1 , AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, Rh10, Rh74, AAV3B, or AAV-2i8, but are different from and not identical to known AAV genes or proteins such as AAV1, AAV2 , AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, Rh10, Rh74, AAV3B or AAV-2i8. In one embodiment, the modified AAV capsid protein/variant AAV capsid protein includes or consists of a sequence that is at least 80%, 85%, 85%, 87%, 88%, 89%, 90%, 91%, 92 %, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, etc. d. up to 99.9% identical to the reference or parent AAV capsid protein, such as AAV (SEQ ID NO: 30-32), LK03 (SEQ ID NO: 33), AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 , AAV9, AAV10, AAV11, AAV12, Rh10, Rh74, AAV3B or AAV-2i8.

[0308] rAAV vectors can be administered to a patient by infusion in a biocompatible vehicle, for example, by intravenous injection. rAAV vectors can be administered alone or in combination with other molecules. Thus, rAAV vectors and other compositions, agents, drugs, biological agents (proteins) can be included in pharmaceutical compositions. Such pharmaceutical compositions can be used, in particular, for administration and delivery to an individual in vivo or ex vivo .

[0309] In certain embodiments, the pharmaceutical compositions also contain a pharmaceutically acceptable carrier or excipient. Such excipients include any pharmaceutical agent that does not itself cause an immune response adverse to the individual to whom the composition is administered, and which can be administered without undue toxicity.

[0310] As used herein, the terms “pharmaceutically acceptable” and “physiologically acceptable” mean a biologically acceptable composition, gaseous, liquid or solid, or mixture thereof, suitable for one or more routes of administration, in vivo delivery or contact. A "pharmaceutically acceptable" or "physiologically acceptable" composition is a material that is not biologically or otherwise undesirable, eg, the material can be administered to a subject without causing significant undesirable biological effects. Thus, such a pharmaceutical composition can be used, for example, in administering a nucleic acid, vector, viral particle or protein to an individual.

[0311] Pharmaceutically acceptable excipients include, but are not limited to, liquids such as water, saline, glycerin, sugars, and ethanol. Pharmaceutically acceptable salts may also be included, for example salts of inorganic acids such as hydrochlorides, hydrobromides, phosphates, sulfates and the like; and salts of organic acids such as acetates, propionates, malonates, benzoates and the like. Excipients also include proteins such as albumin. In addition, such carriers may contain auxiliary agents such as humectants or emulsifiers, pH buffering agents, and the like.

[0312] The pharmaceutical composition may be provided as a salt and may be prepared using many acids including, but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc. Salts tend to be more soluble in aqueous or other protic solvents than the corresponding free base forms. In other cases, the preparation may be a lyophilized powder that may contain any or all of the following: 1-50 mM histidine, 0.1%-2% sucrose and 2-7% mannitol with a pH range of 4.5 to 5.5, which is combined with a buffer before use.

[0313] Pharmaceutical compositions include solvents (aqueous or non-aqueous), solutions (aqueous or non-aqueous), emulsions (eg, oil-in-water or water-in-oil), suspensions, syrups, elixirs, dispersions and suspensions. media, coatings, isotonic and absorption promoting or absorption delaying agents compatible with pharmaceutical administration or in vivo contact or delivery. Aqueous and non-aqueous solvents, solutions and suspensions may include suspending agents and thickeners. Such pharmaceutically acceptable carriers include tablets (coated or uncoated), capsules (hard or soft), microparticles, powders, granules and crystals. The compositions may also include auxiliary active compounds (eg, preservatives, antibacterial, antiviral and antifungal agents).

[0314] Pharmaceutical compositions can be formulated to be compatible with a particular route of administration or delivery, as described herein or known to one of ordinary skill in the art. Thus, pharmaceutical compositions include carriers, diluents or excipients suitable for administration by various routes.

[0315] Compositions suitable for parenteral administration contain aqueous and non-aqueous solutions, suspensions or emulsions of the active compound, where the preparations are generally sterile and may be isotonic with the blood of the intended recipient. Non-limiting illustrative examples include water, buffered saline, Hanks solution, Ringer's solution, dextrose, fructose, ethanol, oils of animal, vegetable or synthetic origin. Aqueous injectable suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran.

[0316] In addition, suspensions of the active compound can be prepared as appropriate oily injectable suspensions. Suitable lipophilic solvents or carriers include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Optionally, the suspension may also contain suitable stabilizers or agents to increase the solubility of the compounds, making it possible to obtain highly concentrated solutions.

[0317] Co-solvents and excipients can be added to the composition. Non-limiting examples of cosolvents contain hydroxyl groups or other polar groups, for example, alcohols such as isopropyl alcohol; glycols such as propylene glycol, polyethylene glycol, polypropylene glycol, glycol ether; glycerol; polyoxyethylene alcohols and polyoxyethylene fatty acid esters. Excipients include, for example, surfactants such as soy lecithin and oleic acid; sorbitan esters such as sorbitan trioleate; and polyvinylpyrrolidone.

[0318] Once the pharmaceutical compositions are received, they can be placed in a suitable container and labeled for treatment. Such notes may include amount, frequency, and route of administration.

[0319] Pharmaceutical compositions and delivery systems suitable for the compositions, methods and uses of the invention are known in the art (see For example, Remington: The Science and Practice of Pharmacy (2003) 20^th ed., Mack Publishing Co., Easton, PA; Remington's Pharmaceutical Sciences (1990) 18^th ed., Mack Publishing Co., Easton, PA; The Merck Index (1996) 12^th ed., Merck Publishing Group, Whitehouse, NJ; Pharmaceutical Principles of Solid Dosage Forms (1993), Technomic Publishing Co., Inc., Lancaster, Pa.; Ansel and Stoklosa, Pharmaceutical Calculations (2001) 11^th ed., Lippincott Williams & Wilkins, Baltimore, MD; and Poznansky et al., Drug Delivery Systems (1980), R. L. Juliano, ed., Oxford, N.Y., pp. 253-315).

[0320] The terms "effective amount" or "sufficient amount" refers to an amount provided in single or multiple doses, alone or in combination with one or more other compositions (therapeutic or immunosuppressive agents, such as a drug), treatments, protocols or means within treatment regimens, a detectable response of any duration (long-term or short-term), an expected or desired response or benefit to an individual of any measurable or detectable extent or of any duration (for example, within minutes, hours, days, months, years or with the achievement cure).

[0321] Dosages may vary and depend on the type, onset, progression, severity, frequency, duration or likelihood of developing the disease being treated, the desired clinical endpoint, prior or concomitant treatment, general health status, age, sex, race or the immunocompetence of the individual and other factors that will be known to those skilled in the art. The amount administered, amount, frequency or duration of dosing may be proportionally increased or decreased as indicated by any undesirable side effects, complications or other risk factors of the treatment or therapy and the status of the individual. Those skilled in the art will be aware of factors that may influence the dosage and timing required to provide an amount sufficient to provide therapeutic or prophylactic benefit.

[0322] The dose to achieve a therapeutic effect, e.g., dose in vector genomes/kilogram body weight (BW/kg), will vary based on several factors including, but not limited to: route of administration, level of heterologous polynucleotide expression required to achieve therapeutic effect, the specific disease being treated, any host immune response to the viral vector, host immune response to the heterologous polynucleotide or expression product (protein), and the stability of the expressed protein. One skilled in the art may determine the dose range of the rAAV/genome vector for treating a patient having a particular disease or disorder, taking into account the above factors as well as other factors.

[0323] Typically, doses will range from at least 1×10 ⁸ vector genomes per kilogram (KG/kg) of the individual's body weight or more, for example, 1×10 ⁹ , 1×10 ¹⁰ , 1×10 ¹¹ , 1 x 10 ¹² , 1 x 10 ¹³ or 1 x 10 ¹⁴ or more vector genomes per kilogram (KG/kg) of an individual's body weight to achieve a therapeutic effect. A dose of rAAV in the range of 1x10 ¹⁰ -1x10 ¹¹ VG/kg in mice and 1x10 ¹² -1x10 ¹³ VG/kg in dogs is effective. Doses may be smaller, for example, the dose may be less than 6×10 ¹² VG/kg. More specifically, the dose may be 5×10 ¹¹ VG/kg or 1×10 ¹² VG/kg.

[0324] Doses of the rAAV vector may be at a level generally at the lower end of the dosage spectrum such that no significant immune response is observed against the heterologous nucleic acid sequence, encoded protein, or inhibitory nucleic acid or rAAV vector. More specifically, the dose may be up to but less than 6 x 10 ¹² VG/kg, such as about 5 x 10 ¹¹ to about 5 x 10 ¹² VG/kg, or more specifically about 5 x 10 ¹¹ VG/kg or about 1×10 ¹² VG/kg.

[0325] As used herein, the term “unit dosage form” refers to physically discrete units formulated as single doses for the individual being treated; each unit contains a predetermined amount, optionally together with a pharmaceutical carrier (excipient, diluent, vehicle or excipient), which when administered in one or more doses is intended to produce the desired effect (eg, prophylactic or therapeutic effect). Unit dosage forms may be, for example, in ampoules and vials, which may include a liquid composition or a lyophilized composition; for example, a sterile liquid carrier may be added prior to administration or in vivo delivery. Individual unit dosage forms may be included in multi-dose kits or containers. rAAV particles and their pharmaceutical compositions can be packaged in single or multiple unit dosage forms for ease of administration and uniformity of dosage.

[0326] Dosages of an "effective amount" or "sufficient amount" for treatment (e.g., to improve or provide a therapeutic benefit or enhancement) are generally effective to provide a response to one, many, or all of the adverse symptoms, effects, or complications of a disease, one or more undesirable symptoms, disorders, diseases, pathologies or complications, such as those caused by or associated with a disease, to a measurable extent, although reducing, inhibiting, suppressing, limiting or controlling the progression or worsening of the disease is a satisfactory outcome.

[0327] An effective amount or sufficient amount may optionally be administered in a single administration, may require multiple administrations, and may optionally be administered alone or in combination with another composition (e.g., agent), treatment, protocol, or regimen treatment. For example, the amount may be increased proportionally depending on the needs of the individual, the type, status and severity of the disease being treated, or the side effects of the treatment (if any). In addition, an effective amount or sufficient amount may not be effective or sufficient when administered in single or multiple doses without a second composition (eg, another drug or agent), treatment, protocol, or treatment regimen because additional doses, amounts or durations of administration beyond such doses, or additional compositions (eg, drugs or agents), treatments, protocols or treatment regimens may be included to be considered effective or sufficient for the specified individual. Amounts considered effective also include amounts resulting in a reduction in the use of another treatment, treatment regimen or protocol, such as the administration of a modified GAA-encoding nucleic acid for the treatment of GAA deficiency (eg, Pompe disease) or other glycogenosis.

[0328] Thus, the methods and uses of the invention also include, but are not limited to, methods and uses that result in a reduced need or use of another compound, agent, drug, treatment regimen, treatment protocol, method or drug. For example, in the case of GAA deficiency, the method or use of the invention is therapeutically beneficial if a less frequently administered or reduced dose is used in the individual, or if the administration of recombinant GAA is eliminated to replenish the individual's deficient or defective GAA. Thus, the invention relates to methods and uses for reducing the need for or use of other treatment or therapy.

[0329] An effective amount or sufficient amount may not be effective for every individual being treated or for the majority of individuals being treated in a given group or population. Effective amount or sufficient amount means effectiveness or sufficiency for a particular individual rather than a group or general population. As is typical with such methods, some individuals will demonstrate a greater response, a lesser response, or no response to a given treatment or use.

[0330] Administration or delivery in vivo to an individual may occur prior to the development of an undesirable symptom, condition, complication, etc., caused by or associated with a disease. For example, screening (eg, genetic) can be used to identify such individuals as candidates for the compositions, methods and uses of the invention. Thus, such individuals include individuals who screen positive for an insufficient amount or deficiency of a functional gene product (eg, GAA or protein deficiency leading to GSD), or who produce an abnormal, partially functional or nonfunctional gene product (eg, GAA or protein involved in GSD).

[0331] Administration or in vivo delivery to an individual in accordance with the methods and uses of the invention as presented herein may be practiced within 1-2, 2-4, 4-12, 12-24, or 24-72 hours after the individual has been identified as having a disease targeted for treatment, having one or more symptoms of the disease, or having been screened and identified as positive as defined herein, even if the individual does not have one or more symptoms of the disease. Of course, the methods and uses of the invention can be practiced 1-7, 7-14, 14-24, 24-48, 48-64 or more days, months or years after the individual has been identified as having the disease targeted for treatment. treatment having one or more disease symptoms or screened and identified as positive, as defined herein.

[0332] The term “improve” means a detectable or measurable improvement in an individual's disease or symptom or underlying cellular response. Detectable or measurable improvement includes subjective or objective reduction, reduction, inhibition, suppression, limitation or control of the occurrence, frequency, severity, progression or duration of a disease or complication caused by or associated with a disease, or improvement of a symptom or underlying cause or effect of a disease or reversal of the disease.

[0333] In the case of Pompe disease, an effective amount will be an amount, for example, that inhibits or reduces the production or accumulation of glycogen, increases or increases the degradation or removal of glycogen. An effective amount will also be an amount that improves eating problems and/or weight gain; poor head and/or neck control; breathing problems and/or lung infections; enlargement and/or thickening of the heart; heart defects; tongue enlargement; problems with swallowing; liver enlargement; low muscle strength; low muscle tone; weakness in the legs, waist and/or arms; dyspnea; problems with exercise; difficulty breathing during sleep; curvature of the spine and/or ankylosis; low muscle tone and/or lack of muscle strength. An effective amount will also be an amount that reduces or inhibits one or more symptoms, or prevents or reduces the progression or worsening of one or more symptoms, or stabilizes one or more symptoms, or improves one or more symptoms in a patient or individual requiring GAA or having Pompe disease.

[0334] Therapeutic dosages will depend, among other factors, on the age and general health of the individual and the severity of the disease or disorder. The therapeutically effective amount in humans will fall within a relatively wide range, which can be determined by medical personnel based on the response of the individual patient.

[0335] Compositions, such as pharmaceutical compositions, can be administered to an individual in such a way as to enable production of the encoded protein. In a specific embodiment, the pharmaceutical compositions contain sufficient genetic material to enable the recipient to produce a therapeutically effective amount of the protein.

[0336] The compositions can be formulated and/or administered in any sterile, biocompatible pharmaceutical carrier, including, but not limited to, saline, buffered saline, dextrose, and water. The compositions can be formulated and/or administered to a patient alone or in combination with other agents (eg, cofactors) that affect hemostasis.

[0337] The methods and uses of the invention include delivery and administration systemically, topically or locally, or by any route, such as by injection or infusion. Delivery of pharmaceutical compositions in vivo can generally be accomplished by injection using a conventional syringe, although other delivery methods are contemplated, such as convection delivery (see, for example, US Pat. No. 5,720,720). For example, the compositions can be administered subcutaneously, epidermally, intradermally, intrathecally, intraorbitally, intramucosally, intranasally, intraperitoneally, intravenously, intrapleurally, intraarterially, intracavitarily, orally, intrahepatically, via the portal vein, or intramuscularly. Other routes of administration include oral and pulmonary administration, suppositories and transdermal administration. A clinician specializing in the treatment of patients with Pompe disease or other glycogen diseases may determine the optimal route for administration of adeno-associated virus vectors based on a number of criteria, including, but not limited to, the patient's condition and the goal of treatment (eg, elevated or decreased GAA levels).

[0338] Songs can be entered individually. In some embodiments, the rAAV particle provides a therapeutic effect without an immunosuppressive agent. The therapeutic effect is optionally maintained for a period of time, for example, 2-4, 4-6, 6-8, 8-10, 10-14, 14-20, 20-25, 25-30 or 30-50 days or more, for example, 50-75, 75-100, 100-150, 150-200 days or more without administration of an immunosuppressive agent. Thus, they provide a therapeutic effect over a period of time.

[0339] The rAAV vectors, methods and uses of the invention can be combined with any compound, agent, drug, treatment or other treatment regimen or protocol having the desired therapeutic, beneficial, additive, synergistic or complementary activity or effect. Examples of combination compositions and treatments include second actives such as biological agents (proteins), agents (eg, immunosuppressive agents), and drugs. Such biological agents (proteins), agents, drugs, treatments and therapies can be administered or carried out before, substantially simultaneously with or after any other method or use of the invention.

[0340] A compound, agent, drug, treatment, or other treatment regimen or protocol can be used as a combination composition or used separately, for example, simultaneously or sequentially (before or after) delivery or administration of a nucleic acid, vector, or rAAV particle. Thus, the present invention relates to combinations in which the method or use of the invention is in combination with any compound, agent, drug, treatment regimen, treatment protocol, treatment method, drug or composition described herein or known to one skilled in the art. this area. The compound, agent, drug, treatment regimen, treatment protocol, method of treatment, drug or composition can be administered or carried out before, substantially simultaneously with or after the administration of the rAAV nucleic acid, vector or particle of the invention to the individual.

[0341] In some embodiments, a nucleic acid, vector, or rAAV particle of the invention is administered to a patient in combination with an immunosuppressive agent or treatment regimen where the patient has an immune response or is at risk of developing an immune response against the rAAV particle and/or GAA protein. Such an immunosuppressive agent or treatment regimen can be used before, substantially simultaneously with, or after administration of the rAAV nucleic acid, vector, or vector of the invention.

[0342] In some embodiments, an individual or patient, such as a human patient, with Pompe disease develops inhibitors to the GAA protein (including anti-GAA antibodies and/or anti-GAA T cells), which may occur after treatment with conventional replacement therapy. enzyme therapy (for example, after administration of recombinantly produced GAA protein). Such GAA inhibitors may appear in patients undergoing enzyme replacement therapy, particularly if the patient has undetectable levels of GAA (as may occur in infantile Pompe disease), causing the patient's immune system to recognize the replacement GAA protein as " alien." In some embodiments, a patient with Pompe disease who has GAA inhibitors is treated with one or more treatment regimens designed to achieve immunological tolerance or reduce the immune response to the GAA protein in the patient prior to, substantially simultaneously with, or subsequent to administration of the rAAV vector of the invention. Such regimens to achieve immunological tolerance or reduce the immune response to the GAA protein may include the administration of one or more immunosuppressive agents, including, but not limited to, methotrexate, rituximab, intravenous gammaglobulin (IVIG), omalizumab and rapamycin in synthetic vaccine particles (SVP™). (rapamycin encapsulated in a biodegradable nanoparticle), and/or the use of one or more methods of immunosuppression, such as B-cell depletion, immunoadsorption and plasmapheresis.

[0343] In some embodiments, the rAAV vector is administered in combination with one or more immunosuppressive agents before, substantially simultaneously with, or after administration of the rAAV vector. In some embodiments, it takes, for example, 1-12, 12-24, or 24-48 hours, or 2-4, 4-6, 6-8, 8-10, 10-14, 14-20, 20-25, 25 -30, 30-50 or more than 50 days after rAAV vector administration. Such administration of immunosuppressive agents is carried out after a period of time after administration of the rAAV vector if a decrease in the encoded protein or inhibitory nucleic acid is observed after initial expression levels over a period of time, for example, 20-25, 25-30, 30-50, 50-75, 75 -100, 100-150, 150-200 or more than 200 days after rAAV vector administration.

[0344] In some embodiments, the immunosuppressive agent is an anti-inflammatory agent. In some embodiments, the immunosuppressive agent is a steroid, such as a corticosteroid. In some embodiments, the immunosuppressive agent is prednisone, prednisolone, cyclosporine (eg, cyclosporine A), mycophenolate, anti-B cell antibody, eg, rituximab; a proteasome inhibitor, such as bortezomib; a mammalian target of rapamycin (mTOR) inhibitor, such as rapamycin; a tyrosine kinase inhibitor, such as ibrutinib; B-cell activating factor (BAFF) inhibitor; or a proliferation-inducing ligand inhibitor (APRIL) or a derivative thereof. In some embodiments, the immunosuppressive agent is an anti-IL-1β agent (e.g., the anti-IL-1β monoclonal antibody canakinumab ( ^Ilaris® )) or an anti-IL-6 agent (e.g., the anti-IL-6 antibody sirukumab or the anti-IL-6 receptor antibody tocilizumab (Actemra ^® )) or a combination thereof.

[0345] Immunosuppressive techniques, including the use of rapamycin, alone or in combination with IL-10, can be used to reduce, reduce, inhibit, prevent, or block humoral and cellular immune responses to the GAA protein. Hepatic gene transfer using the AAV vectors of the invention can be used to induce immunological tolerance to the GAA protein through induction of T regulatory cells ( _Treg ) and other mechanisms. Strategies to reduce (overcome) or avoid the humoral response to AAV during systemic gene transfer include administration of high doses of vector, use of empty AAV capsids as decoys for the adsorption of anti-AAV antibodies, administration of immunosuppressive drugs to reduce, reduce, inhibit, prevent or eradicate humoral immune response to AAV, changing the serotype of the AAV capsid or engineering the AAV capsid so that it is less susceptible to neutralizing antibodies, using plasma exchange cycles to adsorb anti-AAV immunoglobulins and thus reducing the titer of anti-AAV antibodies and using delivery methods such as balloon catheters followed by flushing with saline solution. Such strategies are described in Mingozzi et al ., 2013, Blood, 122:23-36. A review of methods and approaches for inducing GAA tolerance in patients with Pompe disease to improve therapeutic management is given in Doerfler et al ., 2016, Mol. Ther., 3:15053.

[0346] The ratio of empty AAV capsids to rAAV vector may be in the range of or about 100:1-50:1, about 50:1-25:1, about 25:1-10:1, about 10:1-1:1 , approximately 1:1-1:10, approximately 1:10-1:25, approximately 1:25-1:50, or approximately 1:50-1:100. Ratios can also be approximately 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1 or 10:1.

[0347] Amounts of empty AAV capsids to be administered can be calibrated based on the amount (titer) of anti-AAV antibodies produced in a particular individual. Empty AAV capsids can be of any serotype, for example, AAV (SEQ ID NO: 30-32), LK03 (SEQ ID NO: 33), AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, Rh10, Rh74, AAV3B or AAV-2i8.

[0348] Alternatively or additionally, the rAAV vector can be delivered via direct intramuscular injection (eg, into one or more slow-twitch muscle fibers). In another alternative example, a femoral artery catheter can be used to deliver rAAV vectors to the liver via the hepatic artery. Non-surgical methods such as endoscopic retrograde cholangiopancreatography (ERCP) can also be used to deliver rAAV vectors directly to the liver, thereby bypassing the bloodstream and anti-AAV antibodies. Other ductal systems, such as the submandibular gland ducts, can also be used as a portal of entry for delivering rAAV vectors to an individual who has developed or has developed anti-AAV antibodies.

[0349] Additional strategies for reducing humoral immunity to AAV include methods of removing, depleting, capturing and/or inactivating anti-AAV antibodies, commonly referred to as apheresis and, more specifically, plasmapheresis, involving blood products. Apheresis or plasmapheresis is a method in which human plasma is circulated ex vivo (extracorporeal) through a device that modifies the plasma by adding, removing and/or replacing components before returning them to the patient. Plasmapheresis can be used to remove human immunoglobulins (eg, IgG, IgE, IgA, IgD) from a blood product (eg, plasma). This method depletes, captures, inactivates, reduces or removes immunoglobulins (antibodies) that bind to AAV, thereby reducing the titer of anti-AAV antibodies in the treated individual that may be involved in neutralizing the AAV vector. An example is a device consisting of an affinity matrix column for the AAV capsid. Passing a blood product (eg plasma) through an AAV capsid affinity matrix will result in the binding of only anti-AAV antibodies, and in doing so, all isotypes (including IgG, IgM, etc.).

[0350] It is predicted that a sufficient degree of plasmapheresis using an AAV capsid affinity matrix will result in substantially the removal of anti-AAV capsid antibodies and a reduction in anti-AAV capsid antibody titer (load) in humans. In some embodiments, the titer of the individual being treated is reduced to substantially low levels (to <1:5 or less, such as <1:4, or <1:3, or <1:2, or <1 :1). The decrease in antibody titer will be temporary, because B lymphocytes producing anti-AAV capsid antibodies would be expected to gradually cause anti-AAV capsid antibody titers to recover to steady-state levels before plasmapheresis.

[0351] In the event that the titer of pre-existing anti-AAV antibodies was reduced from 1:100 to 1:1, the titer of anti-AAV antibodies was restored by approximately 0.15% (corresponding to a titer of 1:1.2), 0.43% ( 1:1.4), 0.9% (1:1.9), 1.7% (1:2.7) and 3.4% (1:4.4) after 1 hour, 3 hours, 6 hours, 12 hours and 24 hours, respectively, after completion of plasmapheresis. Temporary removal of anti-AAV antibodies from such an individual will correspond to a time window (e.g., approximately 24 hours or less, e.g., 12 hours or less, or 6 hours or less, or 3 hours or less, or 2 hours or less, or 1 hour or less) during which time the AAV vector can be administered to an individual and is predicted to effectively transduce target tissues without significant neutralization of the AAV vector by anti-AAV antibodies.

[0352] In the event that the titer of pre-existing anti-AAV antibodies was reduced from 1:1000 to 1:1, the titer of anti-AAV antibodies was restored by approximately 0.15% (corresponding to a titer of 1:2.5) 0.4% (1 :5.3), 0.9% (1:9.7), 1.7% (1:18) and 3.4% (1:35) at 1 hour, 3 hours, 6 hours, 12 hours and 24 hours, respectively, after completion of plasmapheresis. Thus, the window for introducing the AAV vector will be relatively short.

[0353] Antibodies against AAV may already exist and may be present at levels that reduce or block therapeutic transfer of the GAA gene through vector transduction of target cells. Alternatively, anti-AAV antibodies may occur following exposure to AAV or administration of an AAV vector. If such antibodies arise after administration of the AAV vector, these individuals can also be treated by apheresis, more specifically plasmapheresis.

[0354] In some embodiments, the nucleic acids, expression cassettes, and AAV vectors of the invention may be used in combination with symptomatic and supportive care, including, for example, respiratory support (including mechanical ventilation), physical therapy to strengthen muscles, physical therapy to improve strength and conditioning. opportunities, occupational therapy including the use of canes, walkers and wheelchairs, speech therapy to improve articulation and speech, the use of orthopedic devices including orthotics, and nutritional therapy and feeding tubes to promote proper nutrition and weight gain.

[0355] In some embodiments, the nucleic acids, AAV expression cassettes, and vectors of the invention can be used in combination with pharmacological chaperone therapy (also known as enzyme enhancement therapy), wherein one or more pharmacological chaperones are administered before, simultaneously, or after administration of the nucleic acid expressing AAV cassettes or vectors of the invention for the treatment of GSD such as Pompe disease.

[0356] In some embodiments, the AAV nucleic acids, expression cassettes and vectors of the invention can be used in combination with one or more pharmacological chaperones that can stabilize the GAA protein. Pharmacological chaperones that can be used in combination with the nucleic acids, expression cassettes and AAV vectors of the invention include 1-deoxynojirimycin (1-DNJ, also known as divoglustat), N -butyl-1-deoxynojirimycin (also known as miglustat), N -methyl-DNJ, N-ethyl-DNJ, N-propyl-DNJ, N-pentyl-DNJ, N-hexyl-DNJ, N-heptyl-DNJ, N-octyl-DNJ, N-nonyl-DNJ, N-methylcyclopropyl -DNJ, N-methylcyclopentyl-DNJ, N-2-hydroxyethyl-DNJ, 5-N-carboxypentyl-DNJ and pharmacological chaperones described in US Patent Nos. 6599919 and 9181184 and International Patent Application Publication No. WO/2013/182652.

[0357] In some embodiments, the nucleic acids, AAV expression cassettes and vectors of the invention can be used in combination with adjunctive therapy using one or more β2-agonists, including, for example, clenbuterol, albuterol, formoterol and salmeterol, and as described in the publication international patent application No. WO/2017/049161.

[0358] In certain embodiments, the nucleic acids and expression cassettes of the invention are delivered or administered using AAV vector particles. In other embodiments, the nucleic acids and expression cassettes of the invention can be delivered or administered by other types of viral particles, including retroviral, adenoviral, helper-dependent adenoviral, hybrid adenoviral particles, herpes simplex virus particles, lentiviral particles, poxvirus particles, Epstein virus particles -Barr, vaccinia virus particles and human cytomegalovirus particles.

[0359] In other embodiments, the nucleic acids and expression cassettes of the invention are delivered or administered using a non-viral delivery system. Non-viral delivery systems include, for example, chemical methods, for example, using liposomes, nanoparticles, lipid nanoparticles, polymers, microparticles, microcapsules, micelles or extracellular vesicles, and physical methods, such as gene gun, electroporation, particle bombardment, sonication and magnetofection .

[0360] In some embodiments, the nucleic acids and expression cassettes of the invention are delivered as deproteinized DNA, minicircles, transposons, or blunt-ended linear duplex DNA.

[0361] In other embodiments, the nucleic acids and expression cassettes of the invention are delivered or administered in AAV vector particles or other viral particles that are further encapsulated or complexed with liposomes, nanoparticles, lipid nanoparticles, polymers, microparticles, microcapsules, micelles, or extracellular vesicles .

[0362] The term "lipid nanoparticle" or "LNP" refers to a lipid-based vesicle that can be used to deliver AAV and has dimensions in the nanorange, i.e. from about 10 nm to about 1000 nm, or from about 50 to about 500 nm, or from about 75 to about 127 nm. Without wishing to be bound by theory, it is believed that LNPs provide the nucleic acid expression cassette or AAV vector with partial or complete shielding from the immune system. Shielding allows delivery of the AAV nucleic acid expression cassette or vector into a tissue or cell while avoiding the induction of a significant immune response against the AAV nucleic acid expression cassette or vector in vivo . Shielding may also allow repeated administration without inducing a significant immune response against the nucleic acid expression vector or AAV vector in vivo (eg, in an individual such as a human). Shielding may also improve or increase the efficiency of delivery of an AAV nucleic acid expression cassette or vector in vivo .

[0363] The pI (isoelectric point) of AAV is in the range of about 6 to about 6.5. Thus, the surface of AAV carries a slightly negative charge. In this regard, it may be advantageous if the LNP contains a cationic lipid, such as, for example, an aminolipid. Examples of aminolipids are described in US patent Nos. 9352042, 9220683, 9186325, 9139554, 9126966 9018187, 8999351, 8722082, 8642076, 8569256, 8466122 and 7745651 US publications No. 2016/0213785, 2016/0199485, 2015/0265708, 2014/ 0288146, 2013/0123338, 2013/0116307, 2013/0064894, 2012/0172411 and 2010/0117125.

[0364] The terms “cationic lipid” and “aminolipid” are used interchangeably herein to include lipids and their salts having one, two, three or more fatty acid chains or fatty alkyls and pH-titable amino groups (e.g., alkylamino or dialkylamino groups ). A cationic lipid is typically protonated (ie, positively charged) at a pH below the pKa of the cationic lipid and substantially neutral at a pH above the pKa. Cationic lipids can also be titratable cationic lipids. In some embodiments, the cationic ones contain: a protonatable tertiary amino group (eg, pH-titrated); C18 alkyl chains, wherein each alkyl chain independently has from 0 to 3 (eg, 0, 1, 2 or 3) double bonds; and ether bonds, ester bonds or ketal bonds between the end group and the alkyl chains.

[0365] Cationic lipids may include, but are not limited to, 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLenDMA), 1,2-di-γ -linolenyloxy-N,N-dimethylaminopropane (γ-DLenDMA), 2,2-dilioleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLin-K-C2-DMA, also known as DLin-C2K -DMA, XTC2 and C2K), 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA), dilinoleylmethyl-3-dimethylaminopropionate (DLin-M-C2-DMA, also known as MC2), (6Z,9Z,28Z,31 Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (DLin-M-C3-DMA, also known as MC3), their salts and mixtures. Other cationic lipids also include, but are not limited to, 1,2-distearyloxy-N,N-dimethyl-3-aminopropane (DSDMA), 1,2-dioleyloxy-N,N-dimethyl-3-aminopropane (DODMA), 2 ,2-dilinoleyl-4-(3-dimethylaminopropyl)-[1,3]-dioxolane (DLin-K-C3-DMA), 2,2-dilinoleyl-4-(3-dimethylaminobutyl)-[1,3]- dioxolane (DLin-K-C4-DMA), DLen-C2K-DMA, γ-DLen-C2K-DMA and (DLin-MP-DMA) (also known as 1-B11).

[0366] Additional cationic lipids may include, but are not limited to, 2,2-dilinoleyl-5-dimethylaminomethyl-[1,3]-dioxane (DLin-K6-DMA), 2,2-dilinoleyl-4-N-methylpiperazino -[1,3]-dioxolane (DLin-K-MPZ), 1,2-dilinoleylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2-dilinoleyloxy-3-(dimethylamino)acetoxypropane (DLin-DAC) , 1,2-dilinoleoyloxy-3-morpholinopropane (DLin-MA), 1,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-dilinoleoylthio-3-dimethylaminopropane (DLin-S-DMA), 1-linoleoyl- 2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleoyloxy-3-trimethylaminopropane chloride (DLin-TMA.Cl), 1,2-dilinoleoyl-3-trimethylaminopropane chloride (DLin-TAP.Cl) , 1,2-dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), 3-(N,N-dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-dioleylamino)- 1,2-propanediol (DOAP), 1,2-dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), N,N-dioleyl-N,N-dimethylammonium chloride (DODAC) , N-(1-(2,3-dioleyloxy)propyl)-N, N,N-trimethylammonium chloride (DOTMA), N, N-distearyl-N, N-dimethylammonium bromide (DDAB), N-(1- (2,3-dioleyloxy)propyl)-N, N,N-trimethylammonium (DOTAP), 3-(N-(N',N'-dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), N-(1) bromide ,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethylammonium (DMRIE), 2,3-dioleyloxy-N-[2(spermine-carboxamido)ethyl]-N,N-dimethyl-1-trifluoroacetate propanaminium (DOSPA), dioctadecylamidoglycylspermine (DOGS), 3-dimethylamino-2-(cholest-5-ene-3-beta-hydroxybutane-4-oxy)-1-(cis, cis-9,12-octadecadienoxy)propane (CLinDMA ), 2-[5'-(cholest-5-ene-3-beta-oxy)-3'-oxapentoxy)-3-dimethyl-1-(cis, cis-9',1-2'-octadecadienoxy)propane (CpLinDMA), N,N-dimethyl-3,4-dioleyloxybenzylamine (DMOBA), 1,2-N,N'-dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), 1,2-N,N'-dilinoleylcarbamyl-3- dimethylaminopropane (DLincarbDAP), dexamethasone-spermine (DS) and disubstituted spermine (D2S) or mixtures thereof.

[0367] A number of commercial formulations of cationic lipids can be used, such as LIPOFECTIN® (including DOTMA and DOPE, available from GIBCO/BRL) and LIPOFECTAMINE® (containing DOSPA and DOPE, available from GIBCO/BRL).

[0368] In some embodiments, the cationic lipid may be present in an amount from about 10% by weight of the LNP to about 85% by weight of the lipid nanoparticle, or from about 50% by weight of the LNP to about 75% by weight of the LNP.

[0369] Sterols can impart fluidity to LNP. As used herein, the term “sterol” refers to any naturally occurring sterol of plant (phytosterols) or animal (zoosterols) origin, as well as unnatural synthetic sterols, all of which are characterized by the presence of a hydroxyl group at the 3-position of the steroid A-ring. The sterol may be any sterol commonly used in the art of preparing liposomes, lipid vesicles or lipid particles, most commonly cholesterol. Phytosterols may include campesterol, sitosterol and stigmasterol. Sterols also include sterol-modified lipids, such as those described in US Patent Application Publication No. 2011/0177156. In some embodiments, the sterol may be present in an amount from about 5% by weight of the LNP to about 50% by weight of the lipid nanoparticle, or from about 10% by weight of the LNP to about 25% by weight of the LNP.

[0370] The LNP may contain a neutral lipid. Neutral lipids can contain any type of lipid that exists in an uncharged or neutral zwitterionic form at physiological pH. Such lipids include, but are not limited to, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, dihydrosphingomyelin, cephalin, and cerebrosides. The selection of neutral lipids is usually based on, among other things, particle size and required stability. In some embodiments, the neutral lipid component may be a lipid having two acyl groups (eg, diacylphosphatidylcholine and diacylphosphatidylethanolamine).

[0371] Lipids are available having various acyl groups with varying chain lengths and degrees of saturation, or they can be isolated or synthesized by well known methods. In some embodiments, lipids containing saturated fatty acids with carbon chain lengths ranging from C14 to C22 may be used. In another group of embodiments, lipids with mono- or diunsaturated fatty acids with carbon chain lengths ranging from C14 to C22 are used. In addition, lipids having mixtures of saturated and unsaturated fatty acid chains can be used. Examples of neutral lipids include, but are not limited to, 1,2-dioleoyl-sn-glycero-3-phosphatidyl-ethanolamine (DOPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1-palmitoyl- 2-oleoyl-sn-glycero-3-phosphocholine (POPC) or any related phosphatidylcholine. Neutral lipids may also consist of sphingomyelin, dihydrosphingomyelin, or phospholipids with other end groups such as serine and inositol.

[0372] In some embodiments, the neutral lipid may be present in an amount from about 0.1% by weight of the lipid nanoparticle to about 75% by weight of the LNP, or from about 5% by weight of the LNP to about 15% by weight of the LNP.

[0373] The LNP-encapsulated nucleic acids, expression cassettes, and AAV vector can be included in pharmaceutical compositions, such as a pharmaceutically acceptable carrier or excipient. Such pharmaceutical compositions can be used, in particular, for the administration and delivery of LNP-encapsulated nucleic acids, expression cassettes and AAV vectors to an individual in vivo or ex vivo .

[0374] LNP formulations can be combined with additional components. Non-limiting examples include polyethylene glycol (PEG) and sterols.

[0375] The term "PEG" refers to polyethylene glycol, a linear, water-soluble polymer of repeating PEG units with two terminal hydroxyl groups. PEGs are classified by their molecular weights; for example, PEG 2000 has an average molecular weight of approximately 2000 Daltons, and PEG 5000 has an average molecular weight of approximately 5000 Daltons. PEGs are commercially available from Sigma Chemical Co. and other companies and include, for example, the following functional PEGs: monomethoxy polyethylene glycol (MePEG-OH), monomethoxy polyethylene glycol succinate (MePEG-S), monomethoxy polyethylene glycol succinimidyl succinate (MePEG-S-NHS), monomethoxy polyethylene glycol amine (MePEG-NH2), monomethoxy polyethylene glycol-amine tresylate (MePEG-TRES) and monomethoxypolyethylene glycol imidazolyl carbonyl (MePEG-IM).

[0376] In some embodiments, the PEG may be a polyethylene glycol with an average molecular weight of from about 550 to about 10,000 Daltons and is optionally substituted with an alkyl, alkoxy, acyl, or aryl group. In some embodiments, the PEG may be replaced with methyl at the terminal hydroxyl position. In another preferred embodiment, the PEG may have an average molecular weight of from about 750 to about 5000 Dalton, or from about 1000 to about 5000 Dalton, or from about 1500 to about 3000 Dalton, or about 2000 Dalton, or about 750 Dalton. PEG may be optionally substituted with alkyl, alkoxy, acyl or aryl. In some embodiments, the terminal hydroxyl group can be replaced with a methoxy group or methyl.

[0377] PEG-modified lipids include PEG-dialkyloxypropyl conjugates (PEG-DAA) described in US Pat. Nos. 8,936,942 and 7,803,397. PEG-modified lipids (or lipid-polyoxyethylene conjugates) that can be used may have different anchors. lipid moieties for attaching the PEG moiety to the surface of the lipid vesicle. Examples of suitable PEG-modified lipids include PEG-modified phosphatidylethanolamine and phosphatidic acid, PEG-ceramide conjugates (eg, PEG-CerC14 or PEG-CerC20) described in US Pat. No. 5,820,873, PEG-modified dialkylamines, and PEG-modified 1,2- diacyloxypropane-3-amines. In some embodiments, the PEG-modified lipid may be PEG-modified diacylglycerols and dialkylglycerols. In some embodiments, PEG may be present in an amount from about 0.5% by weight of the LNP to about 20% by weight of the LNP, or from about 5% by weight of the LNP to about 15% by weight of the LNP.

[0378] In addition, the LNP may be PEG-modified and sterol-modified LNP. The LNPs combined with additional components may be the same or separate LNPs. In other words, the same LNP may be PEG-modified and sterol-modified, or, alternatively, the first LNP may be PEG-modified and the second LNP may be sterol-modified. Optionally, the first and second modified LNPs can be combined.

[0379] In some embodiments, prior to encapsulation, the LNPs may have a size ranging from about 10 nm to about 500 nm, or from about 50 nm to about 200 nm, or from 75 nm to about 125 nm. In some embodiments, the LNP in which the nucleic acid, expression vector or AAV vector is encapsulated can have a size ranging from about 10 nm to 500 nm.

[0380] Recombinant cells capable of expressing the GAA sequences of the invention can be used for delivery or administration.

[0381] Deproteinized DNA, such as minicircles and transposons, can be used to introduce or deliver lentiviral vectors. In addition, genome editing technologies such as zinc finger nucleases, meganucleases, TALENs and CRISPR can also be used to deliver the coding sequence of the invention.

[0382] Glycogenosis (GSD) results from the absence of the enzyme that ultimately converts glycogen compounds into glucose. Enzyme deficiency leads to the accumulation of glycogen in tissues. In many cases the defect has systemic consequences, but in some cases the defect is limited to specific tissues. Most patients experience muscle symptoms such as weakness and spasms, although some GSDs manifest as specific syndromes such as hypoglycemic cramps or cardiomegaly.

[0383] The following are non-limiting examples of GSD:

0 - glycogen synthase deficiency; Ia - glucose-6-phosphatase deficiency (Gierke's disease); II - acid maltase deficiency (Pompe disease); III - deficiency of debranching enzyme (Forbes disease); IV - transglucosidase deficiency (Andersen disease, amylopectinosis); V - myophosphorylase deficiency (McArdle disease); VI - phosphorylase deficiency (Hers disease); and VII - phosphofructokinase deficiency (Tarui disease).

[0384] Various forms of GSD affect carbohydrate metabolic pathways. Although at least 14 unique GSDs have been described in the literature, the four that cause clinically significant muscle weakness are Pompe disease (GSD type II, acid maltase deficiency), Forbes disease (GSD type IIIa, debranching enzyme deficiency), McArdle disease (GSD type V, myophosphorylase deficiency) and Tarui disease (GSD type VII, phosphofructokinase deficiency). One form, Gierke's disease (GSD type Ia, glucose-6-phosphatase deficiency), causes clinically significant ischemic organ damage with significant morbidity.

[0385] In general, GSDs are inherited as an autosomal recessive condition. These inherited enzyme defects typically appear in childhood, although some, such as McArdle disease and Pompe disease, have distinct late-onset forms.

[0386] GSD can be treated through enzyme replacement therapy (ERT), for example, using recombinantly produced GAA. Enzyme replacement therapy is an approved treatment for all patients with Pompe disease. It involves intravenous administration of recombinant human acid α-glucosidase. This drug, manufactured by Genzyme, a Sanofi corporation, is Lumizyme (sold as Myozyme outside the United States), and was first approved by the Food and Drug Administration (FDA) in 2006. It is approved for all patients with Pompe disease. The benefit of ERT may be reduced by antibody production, so ERT can also be combined with immune expression.

[0387] GSD can be treated through nutritional therapy, including strict adherence to a diet, can reduce liver size, prevent hypoglycemia, allow reduction of symptoms and growth and development.

[0388] Adjunctive treatment for Pompe disease is symptomatic and supportive. Respiratory support may be required because... most patients have some degree of respiratory distress. Physical therapy may be helpful to strengthen the breathing muscles. Some patients may require artificial respiration via mechanical ventilation (ie, BIPAP or volumetric ventilators) during the night and/or periods of the day. Additionally, additional support may be needed during respiratory tract infections. Mechanical ventilation can be performed using non-invasive or invasive methods. It is best that the decision about the duration of respiratory support is made by the family in careful consultation with the patient's physicians and other members of the health care team, depending on the patient's specific needs. In non-infantile Pompe disease, a high-protein diet may be beneficial.

[0389] Physical therapy is recommended to improve strength and physical performance. Occupational therapy, including the use of a cane or walker, may be required. Ultimately, some individuals may need to use a wheelchair. Speech therapy may be helpful in improving articulation and speech in some patients.

[0390] Orthopedic devices, including orthotics, may be recommended for some patients. Some orthopedic symptoms, such as contractures or spinal deformities, may require surgery.

[0391] Because Pompe disease weakens the muscles used for chewing and swallowing and may require measures to ensure proper nutrition and weight gain. Some patients may require a specialized, high-calorie diet and training in how to change the size and texture of food to reduce the risk of aspiration. Some infants may need a feeding tube passed through the nose, then through the esophagus into the stomach (nasogastric tube). For some children, a feeding tube may need to be inserted directly into the stomach through a small surgical opening in the abdominal wall. Some individuals with late-onset Pompe disease may require a soft food diet, but some require feeding tubes.

[0392] Individuals can be tested for one or more liver enzymes in the event of an undesirable response to treatment or to determine whether such individuals are suitable for treatment with the method of the invention, prior to treatment. Thus, candidate individuals can be screened for amounts of one or more liver enzymes before or after treatment with the method of the invention. Treated individuals may be monitored post-treatment for elevated liver enzymes periodically, for example, every 1-4 weeks, 1-6 months, 6-12 months, or 1, 2, 3, 4, 5 or more years.

[0393] Examples of liver enzymes include alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH), but other enzymes that are indicators of liver damage may also be monitored. The normal level of these enzymes in the bloodstream is generally defined as a range having an upper level above which the enzyme level is considered elevated and thus an indicator of liver damage. The normal range depends in part on the standards used by the clinical laboratory performing the test.

[0394] The invention relates to kits with packaging material and one or more components therein. The kit typically includes a label or package insert that includes a description of the components or instructions for the in vitro , in vivo , or ex vivo use of the components therein. The kit may contain a group of such components, for example, a rAAV particle and, optionally, a second active agent, such as another compound, agent, drug or composition.

[0395] The term "kit" refers to a physical structure that houses one or more components of a kit. The packaging material can keep components sterile and can be made from a material commonly used for such purposes (eg paper, corrugated cardboard, glass, plastic, foil, ampoules, vials, tubes, etc.).

[0396] Labels or inserts may include identifying information about one or more components, dosages, clinical pharmacology of the active ingredients, including mechanisms of action, pharmacokinetics, and pharmacodynamics. Labels or inserts may include information about the manufacturer, lot numbers, place and date of production, and expiration date. Labels or inserts may include information about the manufacturer, lot numbers, location and date of production. Labels or inserts may include information about the disease for which the kit component may be used. The labels or inserts may include instructions for a clinician or individual to use one or more of the components of the kit in a method, application, treatment protocol, or treatment regimen. The instructions may include dosage, frequency or duration of administration, and instructions for the practice of any of the methods, uses, treatment protocols, or prophylactic or treatment regimens provided herein.

[0397] Labels or inserts may include information about any benefit that the component may provide, such as a prophylactic or therapeutic benefit. Labels or package inserts may include information about potential unwanted side effects, complications, or reactions, such as a warning to the individual or clinician regarding situations where use of a particular composition would be inappropriate. Undesirable side effects or complications may also occur when the individual has taken, will be taking, or is currently taking one or more other drugs that may be incompatible with the composition, or the individual has been, will be, or is currently undergoing another treatment protocol or regimen. treatments that would be incompatible with the composition, and thus the instructions may include information about such incompatibility.

[0398] Labels or inserts include “printed materials,” such as paper or cardboard, either separate from or affixed to a component, kit, or packaging material (e.g., a box), or affixed to a vial, tube, or vial constituting a component of the kit. The labels or inserts may further include a machine-readable medium such as a printed barcode label, a disk, an optical disk such as a CD or DVD-ROM/RAM, DVD, MP3, magnetic tape, or electrical storage devices such as RAM and ROM, or their hybrids, such as magnetic/optical media, FLASH media or memory cards.

[0399] Unless otherwise specified, all technical and scientific terms used in this description have the meaning commonly understood by one skilled in the art to which the present invention belongs. Although methods and materials similar or equivalent to those presented herein may be used in the practice or testing of the present invention, suitable methods and materials are provided herein.

[0400] All patents, patent applications, publications and other references, GenBank entries and ATCC entries cited herein are incorporated by reference in their entirety. In the event of a conflict, this description, including the definition, will control.

[0401] All features presented in the present description can be combined in any combination. Each feature presented herein may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless specifically stated otherwise, the described features (e.g., modified GAA-encoding nucleic acids, expression cassettes containing modified GAA-encoding nucleic acids, and rAAV particles containing modified GAA-encoding nucleic acids) are examples of the same or similar signs.

[0402] As used herein, terms in the singular include references to the plural unless the context clearly indicates otherwise. Thus, for example, a reference to a “nucleic acid” includes a plurality of such nucleic acids, a reference to a “vector” includes a plurality of such vectors, and a reference to a “virus” or “particle” includes a plurality of such viruses/particles.

[0403] For the purposes of the invention, all numerical values or numerical ranges include integers in such ranges and fractions of values or integers in such ranges unless the context clearly indicates otherwise. Thus, for example, a reference to 86% or more of identity includes 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, etc., as well as 86.1%, 86.2%, 86.3%, 86.4%, 86.5%, etc., 87.1 %, 88.2%, 88.3%, 88.4%, 88.5%, etc.

[0404] A reference to an integer with "more than" or "less than" includes any number greater than or less than the reference number, respectively. Thus, for example, a reference to less than 127, 126, 125, 124, 123, 122, 121, 120, 119, 118, 117, 116, 115, 114, 113, 112, 111, 110, etc. down to zero (0); and "less than 10" includes 9, 8, 7, etc. down to zero (0).

[0405] As used herein, all numerical values or ranges include subranges and fractions of values and integers within such ranges and subranges and the wrong 1, as well as the file okay thanks fractions of integers within such ranges, unless the context clearly indicates otherwise. Thus, as an example, reference to a numeric range such as 1-10 includes 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9 , 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 3-4, 3-5, 3-6, 3-7, 3 -8, 3-9, 3-10, etc.; and 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, as well as 1.1, 1.2, 1.3, 1.4, 1.5, etc. Thus, reference to the range 1-50 includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 and etc. up to and including 50, as well as 1.1, 1.2, 1.3, 1.4, 1.5, etc., 2.1, 2.2, 2.3, 2.4, 2 ,5, etc.

[0406] A reference to a series of ranges includes ranges that combine the values of the boundaries of different ranges in the series. So, as an example, reference to a series of ranges such as 1-10, 10-20, 20-30, 30-40, 40-50, 50-60, 60-75, 75-100, 100-150, 150-200, 200-250, 250-300, 300-400, 400-500, 500-750, 750-850, includes ranges 1-20, 1-30, 1-40, 1-50, 1-60, 10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90, 50- 75, 50-100, 50-150, 50-200, 50-250, 100-200, 100-250, 100-300, 100-350, 100-400, 100-500, 150-250, 150-300, 150-350, 150-400, 150-450, 150-500, etc.

[0407] The present invention is generally described herein by using affirmative terms to describe numerous embodiments and aspects. The present invention also specifically includes embodiments in which a particular subject matter of the invention is excluded, in whole or in part, such as substances or materials, process steps and conditions, methods. For example, in some embodiments or aspects of the invention, materials and/or method steps are eliminated. Thus, although the present invention is not generally expressed herein in terms of what the invention does not include, aspects that are not specifically excluded are nevertheless presented herein.

[0408] A number of embodiments of the invention are described. However, one skilled in the art may make various changes and modifications to the invention to adapt it to different uses and conditions without departing from the spirit and scope of the invention. Accordingly, the following examples are intended to illustrate and not to limit the scope of the invention claimed in any way.

EXAMPLE 1

Table 1 - Overview of GAA Expression Cassette

Expression Cassette (SEQ ID NO: 16-24) SEQ ID NO Codon-optimized version PolyA signal BGH Vector Packaging (SEQ ID NO: 30-32) pAAV-ApoE/hAAT.fixUTR.GAA13.BGH 16 GAA13 With reduced CpG content SPK-AAV-01 pAAV-ApoE/hAAT.GAA13.BGH 17 GAA13 With reduced CpG content SPK-AAV-02 pAAV-ApoE/hAAT.GAA7.BGH 18 GAA7 With reduced CpG content SPK-AAV-03 pAAV-ApoE/hAAT.GAA8.BGH 19 GAA8 With reduced CpG content SPK-AAV-04 pAAV-ApoE/hAAT.GAA2.wtBGH 20 GAA2 Wild type BGH SPK-AAV-05 pAAV-ApoE/hAAT.GAA5.wtBGH 21 GAA5 Wild type BGH SPK-AAV-06 pAAV-ApoE/hAAT.GAA7.wtBGH 22 GAA7 Wild type BGH SPK-AAV-07 pAAV-ApoE/hAAT.GAA8.wtBGH 23 GAA8 Wild type BGH SPK-AAV-08 pAAV-ApoE/hAAT.GAA13.wtBGH 24 GAA13 Wild type BGH SPK-AAV-09 pAAV-ApoE/hAAT.GAA13.BGH 17 GAA13 With reduced CpG content SPK-AAV-10*

*SPK-AAV-10 is packaged in the AAV6 capsid, and all others are packaged in the AAV-4-1 capsid variant described in International Patent Application Publication No. WO 2016/210170.

[0409] GAA expression cassettes are shown in Figures 1-9. All contain 5' and 3' flanking AAV inverted terminal repeats (ITRs), a liver-specific ApoE/hAAT enhancer/promoter sequence functionally linked to an optimized GAA coding sequence, including the human hemoglobin beta subunit intron (HBB2) followed by wild-type or CpG-reduced bovine growth hormone (bGH) polyadenylation (poly-A) sequence.

SEQ ID NO: 1:Subsequence GAA2 nucleic acid

ATGGCTTTCCTGTGGCTGCTGAGCTGCTGGGCTCTGCTGGGCACCACCTTTGGGCTGCTGGTGCCTAGGGAGCTGTCTGGGTCTAGCCCTGTGCTGGAGGAGACTCACCCTGCCCATCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCTCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCCACCCAGTGTGATGTGCCCCCCAACAGCAGGTTTGACTGTGCCCCTGACAAGGCCATTACCCAGGAGCAGTGTGAGGCCAGGGGCTGCTGCTACATTCCAGCTAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTATCCTAGCTATAAACTGGAGAACCTGAGCAGCTCTGAGATGGGCTATACTGCCACCCTGACTAGGACTACTCCCACCTTTTTTCCTAAGGATATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCACTTCACTATTAAGGACCCTGCCAATAGGAGGTATGAAGTGCCTCTGGAGACTCCTCATGTGCACTCTAGGGCCCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTGCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGAGCACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCATCTGAGCCCTCTGATGCTGAGCACCTCTTGGACCAGGATCACCCTGTGGAATAGGGATCTGGCCCCCACCCCTGGGGCTAATCTGTATGGCTCTCATCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTTCTGCTGAACAGCAATGCCATGGATGTGGTGCTGCAGCCCTCTCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTGTACATCTTCCTGGGCCCTGAGCCCAAGTCTGTGGTCCAGCAGTATCTGGATGTGGTGGGCTACCCCTTTATGCCCCCCTATTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGGTATTCTTCTACTGCTATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCTCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTATATGGACTCTAGGAGGGATTTCACCTTCAACAAGGATGGCTTCAGGGACTTCCCTGCTATGGTCCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCCATCAGCAGCTCTGGCCCTGCTGGCAGCTATAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTTATCACTAATGAAACTGGGCAGCCCCTGATTGGCAAGGTGTGGCCTGGCTCTACTGCCTTCCCTGACTTCACCAACCCCACTGCTCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGACCAGGTGCCTTTTGATGGCATGTGGATTGACATGAATGAGCCCAGCAACTTCATCAGGGGCTCTGAGGATGGGTGCCCCAATAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACCCTGCAGGCTGCCACTATTTGTGCCAGCTCTCACCAGTTCCTGAGCACCCACTACAACCTGCACAATCTGTATGGCCTGACTGAGGCCATTGCCAGCCACAGGGCCCTGGTGAAGGCCAGGGGCACTAGGCCCTTTGTGATCTCTAGAAGCACCTTTGCTGGCCATGGGAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCCCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAACACCTCTGAAGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTCTACCCTTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCTCAGGAGCCTTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCTCTGCTGCCCCACCTGTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTCCTGGAGTTTCCTAAGGATAGCAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCCCTGCTGATTACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACTTGGTATGACCTGCAGACTGTGCCTGTGGAAGCCCTGGGCAGCCTGCCTCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCCCTGGACACCATTAATGTGCATCTGAGGGCTGGGTATATTATCCCCCTGCAGGGGCCTGGGCTGACTACCACTGAGAGCAGGCAGCAGCCTATGGCCCTGGCTGTGGCTCTGACTAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTACACCCAGGTGATTTTCCTGGCCAGGAACAACACCATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAAGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACAGCCCTGACACCAAGGTGCTGGATATTTGTGTGAGCCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGA

SEQ ID NO: 2: GAA5 Nucleic Acid Sequence

ATGGCTTTCCTGTGGCTGCTGTCTTGCTGGGCCCTGCTGGGGACTACCTTTGGCCTGCTGGTGCCCAGGGAACTGTCTGGCTCTAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCTTCTAGGCCTGGCCCCAGGGATGCCCAGGCCCACCCTGGCAGGCCAAGGGCTGTGCCCACCCAGTGTGATGTGCCCCCCAACTCTAGATTTGATTGTGCCCCTGATAAGGCCAT CACCCAGGAGCAGTGTGAGGCTAGGGGCTGCTGCTACATCCCTGCTAAGCAGGGCCTGCAGGGGGCTCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTATCCCTCTTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCCACCCTGACCAGGACTACTCCCACCTTCTTCCCCAAGGACATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCATT TCACCATCAAGGATCCTGCCAACAGGAGGTATGAGGTGCCTCTGGAGACCCCCATGTGCACAGCAGGGCTCCTTCTCCCCTGTACTCTGTGGAGTTCTCTGAGGAACCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTCCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGATCAGTTCCTGCAGCTGTCCACTTCTCTGCCTAGCCAGTACATCACTGGGCT GGCTGAGCACCTGAGCCCTCTGATGCTGAGCACCTCTTGGACTAGGATCACCCTGTGGAACAGGGACCTGGCCCCCACCCCTGGGGCCAACCTGTATGGCAGCCACCCCTTCTATCTGGCCCT GGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAATAGCAATGCTATGGATGTGGTGCTGCAGCCCAGCCCTGCCCTGTCTTGGAGGAGCACTGGGGGCATCCTGGATGTGTACATTTTC CTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTACCCCTTCATGCCTCCCTACTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGCTACAGCTCTACTGCCATC ACCAGGCAGGTGGTGGAATATGACCAGGGCCCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGACTCTAGGAGGACTTCACCTTCAATAAGGATGGCTTCAGAGACTTC CCTGCCATGGTGCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCCATCAGCTCTTCTGGCCCTGCTGGCTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGGCAGCCCCTGATTGGGAAGGTGTGGCCTGGCTCTACTGCCTTCCCTGACTTCACCAATCCTACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTC CATGACCAGGTGCCCTTTGATGCATGTGGATTGACATGAATGAGCCCTCTAATTTCATCAGGGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCTATGTGCCTGGGGT GGTGGGGGGCACCCTGCAGGCTGCCACCATCTGTGCTAGCTCTCACCAGTTCCTGAGCACCCACTACAATCTGCATAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCACAGGGCCCTG GAAGGCTAGGGGCACCAGGCCCTTTGTGATTTCTAGGAGCACTTTTGCTGGCCATGGCAGGTATGCTGGGCACTGGACTGGGGATGTGTGGTCTAGCTGGGAGCAGCTGGCTTCTTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGGGTTCCTGGGCAACACTTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTCTACC CTTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCCCAGGAGCCCTACAGCTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCACCT GTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCTAGGCCTCTGTTCCTGGAGTTCCCCAAGGACTCTAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCC CTGCTGATCACTCCTGTGCTGCAGGCTGGGAAGGCTGAGGTGACTGGCTATTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCCCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTGGATACCATCAATGTGCACCTGAGGGCTGGCTACATCATTCCCCTGCAGGGCCCTGGCCTGA CCACTACTGAGTCTAGGCAGCAGCCCATGGCCCTGGCTGTGGCCCTGACCAAGGGGGGGGAGGCTAGGGGGGAGCTGTTTTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTACACTCAGGTGATCTTCCTGGCCAGGAACAATACCATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCC CCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTATAGCCCTGATACCAAGGTGCTGGATATTTGTGTGAGCCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGA

SEQ ID NO: 3: GAA7 Nucleic Acid Sequence

ATGGCTTTCCTGTGGCTGCTGTCTTGTTGGGCTCTGCTGGGCACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGCAGCAGCCCTGTGCTGGAGGAGACCCACCCTGCTCATCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCCCAGGCTCACCCTGGGAGACCCAGGGCTGTGCCCACTCAGTGTGATGTGCCCCCCAACAGCAGGTTTGACTGTGCTCCTGACAAGGCTATCACCCAGGAGCAGTGTGAGGCCAGGGGGTGCTGCTACATTCCTGCTAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCTCTTATCCCAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGCTACACTGCCACCCTGACCAGGACCACTCCCACCTTCTTTCCCAAGGATATTCTGACTCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCACTTCACTATCAAGGACCCTGCCAATAGGAGGTATGAGGTGCCCCTGGAGACTCCTCATGTGCATAGCAGGGCCCCTTCTCCTCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTGCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGAGCACTTCTCTGCCCAGCCAGTACATTACTGGGCTGGCTGAGCATCTGAGCCCCCTGATGCTGAGCACCTCTTGGACCAGGATCACCCTGTGGAACAGGGACCTGGCCCCCACTCCTGGGGCTAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTTCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCTCTCCAGCCCTGTCTTGGAGGAGCACTGGGGGCATTCTGGATGTGTACATTTTCCTGGGGCCTGAACCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTACCCCTTCATGCCCCCCTATTGGGGGCTGGGGTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCATTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGATAGCAGGAGGGATTTCACCTTCAACAAGGATGGCTTCAGGGACTTTCCTGCCATGGTGCAGGAGCTGCACCAGGGGGGCAGGAGGTATATGATGATTGTGGACCCTGCTATCAGCAGCTCTGGCCCTGCTGGCTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTTATCACTAATGAAACTGGCCAGCCTCTGATTGGCAAGGTCTGGCCTGGCTCTACTGCCTTCCCTGATTTTACTAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGATCAGGTGCCTTTTGATGGCATGTGGATTGATATGAATGAACCAAGCAACTTCATCAGAGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACCATTTGTGCTAGCAGCCACCAGTTCCTGAGCACCCACTACAATCTGCACAACCTGTATGGCCTGACTGAAGCCATTGCCAGCCATAGGGCCCTGGTGAAGGCCAGGGGCACTAGGCCTTTTGTGATCAGCAGGAGCACTTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCAGCTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATTCTGCAGTTTAACCTGCTGGGGGTGCCCCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTTTATCCCTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCTCAGGAGCCCTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCACCTGTATACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTCCTGGAGTTCCCCAAGGACAGCAGCACCTGGACTGTGGATCATCAGCTGCTGTGGGGGGAGGCCCTGCTGATCACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTCACTGGCTACTTCCCTCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCTCTGGGCAGCCTGCCCCCCCCCCCTGCTGCTCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCTCCCCTGGACACCATCAATGTGCACCTGAGGGCTGGCTACATTATCCCCCTGCAGGGCCCAGGGCTGACTACCACTGAGAGCAGACAGCAGCCCATGGCTCTGGCTGTGGCCCTGACCAAGGGGGGGGAAGCTAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTATACCCAGGTGATCTTCCTGGCTAGGAACAACACCATTGTCAATGAGCTGGTGAGGGTGACTTCTGAGGGGGCTGGGCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCTCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACAGCCCTGACACCAAGGTGCTGGACATCTGTGTGTCTCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGA

SEQ ID NO: 4: GAA8 Nucleic Acid Sequence

ATGGCCTTCCTGTGGCTGCTGTCTTGCTGGGCTCTGCTGGGGACCACCTTTGGCCTGCTGGTCCCCAGGGAGCTGTCTGGCTCTTCTCCTGTCCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCCACCCAGTGTGATGTGCCTCCCCAACAGCAGGTTTGACTGTGCCCCTGACAAGGC CATCACCCAGGAGCAGTGTGAGGCCAGGGGCTGCTGCTATATCCCTGCCAAGCAGGGCCTGCAGGGGGCTCAGATGGGCCAGCCCTGGTGCTTCTTTCCCCCCTCTTATCCTAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGGTACACTGCCACCCTGACCAGGACCACCCCCACTTTCTTCCCTAAGGACATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAATAGGCTGCACT TTACTATCAAGGACCCTGCCAACAGGAGGTATGAGGTGCCTCTGGAGACCCCCATGTGCATTCTAGGGCCCCCAGCCCCCTGTACTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGACAGCTGGATGGCAGGGTCCTGCTGAACACCACTGTGGCTCCCCTGTTTTTTGCTGACCAGTTCCTGCAGCTGAGCACCAGCCTGCCCAGCCAGTACATCACTGGGCTG GCTGAGCACCTGAGCCCCCTGATGCTGAGCACCAGCTGGACCAGGATCACCCTGTGGAACAGGGATCTGGCTCCTACCCCTGGGGCCAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTG GAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCAGCCCTGCCCTGAGCTGGAGGTCTACTGGGGGCATCCTGGATGTGTACATCTTT CTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTATCCTTTTATGCCCCCCTATTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCAT CACCAGACAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGACAGCAGGAGGGACTTCACCTTTAACAAGGATGGCTTTAGACT TCCCTGCCATGGTGCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCAGCCATCAGCAGCTCTGGGCCTGCTGGGTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGGAGCACTGCCTTCCCTGATTTTACCAACCCCACTGCCCTGGCCTGGTGGGAGGATATGGTGGCTGAG TTTCATGACCAGGTGCCCTTTGATGCATGTGGATTGACATGAATGAGCCCAGCAATTTCATCAGGGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAATCCTCCCTATGTGCCTGG GGTGGTGGGGGGCACCCTGCAGGCTGCCACCATCTGTGCCTCTAGCCACCAGTTCCTGAGCACCCACTATAACCTGCATAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGAGCCCT GGTGAAGGCCAGAGGGACCAGGCCCTTTGTGATCTCTAGGAGCACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCTCTTGGGAGCAGCTGGCCAGCTCTGTGCCAGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAATACCTCTGAAGAGCTGTGTGTGAGGTGGACTCAGCTGGGGGCCT TCTATCCCTTCATGAGGAACCACAACAGCCTGCTGTCTCTGCCCCAGGAGCCCTACAGCTTCTCTGAGCCTGCTCAGCAGGCTATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCC CCATCTGTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTTCTGGAGTTTCCCAAGGACAGCAGCACCTGGACTGTGGACCATCAGCTGCTGTGGGGG A GGGCTGACCACCACTGAGAGCAGGCAGCAGCCCATGGCCCTGGCTGTGGCCCTGACTAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGA GAGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAACACCATTGTGAATGAGCTGGTGAGGGTGACTTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCC ACTGCCCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGTCTAACTTCACCTACAGCCCTGATACTAAGGTGCTGGATATCTGTGTGAGCCTGCTGATGGGGGAGCAGTTTCTGGTGAGCTGGTGCTGA

SEQ ID NO: 5: GAA13 Nucleic Acid Sequence

ATGGCCTTTCTGTGGCTGCTGTCCTGCTGGGCCCTGCTGGGGACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGGAGCAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCCAGCAGGCCTGGCCCTAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCTACCCAGTGTGATGTGCCACCCAATTCTAGGTTTGACTGTGCTCCTGACAAGGC CATCACTCAGGAGCAGTGTGAAGCTAGGGGGTGCTGCTACATCCCAGCCAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTACCCTAGCTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCTACCCTGACCAGGACCACTCCTACCTTCTTCCCCAAGGACATCCTGACTCTGAGGCTGGATGTCATGATGGAGACTGAAAATAGGCTGCACT TCACCATCAAGGACCCTGCCAATAGGAGGTATGAGGTGCCTCTGGAGACCCCCATGTGCATAGCAGGGCTCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTCATTGTGAGGAGACAGCTGGATGGGAGGGTGCTGCTGAACACTACTGTGGCTCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGTCTACCAGCCTGCCCAGCCAGTACATCACTGGGCTG GCTGAGCATCTGAGCCCCCTGATGCTGAGCACCAGCTGGACCAGGATCACTCTGTGGAACAGGGATCTGGCCCCCACTCCTGGGGCCAACCTGTATGGGAGCCATCCCTTCTACCTGGCCCTG GAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCCATGGATGTGGTGCTGCAGCCTAGCCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTCTACATCTTC CTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTATCTGGATGTGGTGGGGTATCCCTTCATGCCCCCCTACTGGGGCCTGGGCTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCAT CACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCTCTGGATGTGCAGTGGAATGACCTGGACTATATGGATTCTAGGAGAGACTTTACTTTTAACAAGGATGGCTTCAGGGATTT CCCTGCCATGGTGCAGGAGCTGCACCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCTATTAGCAGCTCTGGCCCTGCTGGGTCTTACAGGCCTTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGCAGCACTGCCTTCCCTGACTTCACCAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGT TCCATGACCAGGTGCCCTTTGATGGGATGTGGATTGACATGAATGAGCCCTCTAACTTCATCAGGGGGTCTGAGGATGGCTGCCCCAAATGAGCTGGAGAACCCCCTATGTGCCTGGGG TGGTGGGGGGCACTCTGCAGGCTGCCACTATCTGTGCTTCTTCTCACCAGTTTCTGAGCACCCACTATAATCTGCACAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGGGCCCTG GAAGGCCAGGGGCACCAGGCCCTTTGTGATCAGCAGGTCTACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGTCTTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGGCTTTCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTT A GGCCCTGCTGATCACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCTCCCCCC CCTGCTGCCCCCAGGGAGCCTGCCATTCATTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGACACCATCAATGTGCACCTGAGGGCTGGGTACATCATCCCCCTGCAGGGCCCTCC TGACCACCACTGAGAGCAGGCAGCAGCCCATGGCCCTGGCTGTGGCTCTGACCAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGTCTCTGGAGGTGCTGGAGAGGGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAATACTATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACT GCCCCCCAGCAGGTCCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACTCTCCTGACACCAAGGTGCTGGACATTTGTGTGTCTCTGCTGATGGGGGAGCAGTTCCCTGGTGAGCTGGTGCTGA

SEQ ID NO: 6: Poly-A GAA2 Nucleic Acid Sequence BGH WT

ATGGCTTTCCTGTGGCTGCTGAGCTGCTGGGCTCTGCTGGGCACCACCTTTGGGCTGCTGGTGCCTAGGGAGCTGTCTGGGTCTAGCCCTGTGCTGGAGGAGACTCACCCTGCCCATCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCTCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCCACCCAGTGTGATGTGCCCCCCAACAGCAGGTTTGACTGTGCCCCTGACAAGGCCATTACCCAGGAGCAGTGTGAGGCCAGGGGCTGCTGCTACATTCCAGCTAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTATCCTAGCTATAAACTGGAGAACCTGAGCAGCTCTGAGATGGGCTATACTGCCACCCTGACTAGGACTACTCCCACCTTTTTTCCTAAGGATATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCACTTCACTATTAAGGACCCTGCCAATAGGAGGTATGAAGTGCCTCTGGAGACTCCTCATGTGCACTCTAGGGCCCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTGCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGAGCACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCATCTGAGCCCTCTGATGCTGAGCACCTCTTGGACCAGGATCACCCTGTGGAATAGGGATCTGGCCCCCACCCCTGGGGCTAATCTGTATGGCTCTCATCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTTCTGCTGAACAGCAATGCCATGGATGTGGTGCTGCAGCCCTCTCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTGTACATCTTCCTGGGCCCTGAGCCCAAGTCTGTGGTCCAGCAGTATCTGGATGTGGTGGGCTACCCCTTTATGCCCCCCTATTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGGTATTCTTCTACTGCTATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCTCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTATATGGACTCTAGGAGGGATTTCACCTTCAACAAGGATGGCTTCAGGGACTTCCCTGCTATGGTCCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCCATCAGCAGCTCTGGCCCTGCTGGCAGCTATAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTTATCACTAATGAAACTGGGCAGCCCCTGATTGGCAAGGTGTGGCCTGGCTCTACTGCCTTCCCTGACTTCACCAACCCCACTGCTCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGACCAGGTGCCTTTTGATGGCATGTGGATTGACATGAATGAGCCCAGCAACTTCATCAGGGGCTCTGAGGATGGGTGCCCCAATAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACCCTGCAGGCTGCCACTATTTGTGCCAGCTCTCACCAGTTCCTGAGCACCCACTACAACCTGCACAATCTGTATGGCCTGACTGAGGCCATTGCCAGCCACAGGGCCCTGGTGAAGGCCAGGGGCACTAGGCCCTTTGTGATCTCTAGAAGCACCTTTGCTGGCCATGGGAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCCCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAACACCTCTGAAGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTCTACCCTTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCTCAGGAGCCTTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCTCTGCTGCCCCACCTGTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTCCTGGAGTTTCCTAAGGATAGCAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCCCTGCTGATTACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACTTGGTATGACCTGCAGACTGTGCCTGTGGAAGCCCTGGGCAGCCTGCCTCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCCCTGGACACCATTAATGTGCATCTGAGGGCTGGGTATATTATCCCCCTGCAGGGGCCTGGGCTGACTACCACTGAGAGCAGGCAGCAGCCTATGGCCCTGGCTGTGGCTCTGACTAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTACACCCAGGTGATTTTCCTGGCCAGGAACAACACCATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAAGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACAGCCCTGACACCAAGGTGCTGGATATTTGTGTGAGCCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGACTCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGA

SEQ ID NO: 7: Poly-A BGH WT GAA5 Nucleic Acid Sequence

ATGGCTTTCCTGTGGCTGCTGTCTTGCTGGGCCCTGCTGGGGACTACCTTTGGCCTGCTGGTGCCCAGGGAACTGTCTGGCTCTAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCTTCTAGGCCTGGCCCCAGGGATGCCCAGGCCCACCCTGGCAGGCCAAGGGCTGTGCCCACCCAGTGTGATGTGCCCCCCAACTCTAGATTTGATTGTGCCCCTGATAAGGCCATCA CCCAGGAGCAGTGTGAGGCTAGGGGCTGCTGCTACATCCCTGCTAAGCAGGGCCTGCAGGGGGCTCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTATCCCTCTTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCCACCCTGACCAGGACTACTCCCACCTTCTTCCCCAAGGACATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCATTTC ACCATCAAGGATCCTGCCAACAGGAGGTATGAGGTGCCTCTGGAGACCCCCCATGTGCACAGCAGGGCTCCTTCTCCCCTGTACTCTGTGGAGTTCTCTGAGGAACCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTCCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGATCAGTTCCTGCAGCTGTCCACTTCTCTGCCTAGCCAGTACATCACTGGGCTGGCT GAGCACCTGAGCCCTCTGATGCTGAGCACCTCTTGGACTAGGATCACCCTGTGGAACAGGGACCTGGCCCCCACCCCTGGGGCCAACCTGTATGGCAGCCACCCCTTCTATCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAATAGCAATGCTATGGATGTGGTGCTGCAGCCCAGCCCTGCCCTGTCTTGGAGGAGCACTGGGGGCATCCTGGATGTGTACATTTTC CTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTACCCCTTCATGCCTCCCTACTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGCTACAGCTCTACTGCCATC ACCAGGCAGGTGGTGGAATATGACCAGGGCCCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGACTCTAGGAGGACTTCACCTTCAATAAGGATGGCTTCAGAGACTTC CCTGCCATGGTGCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCCATCAGCTCTTCTGGCCCTGCTGGCTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGGCAGCCCCTGATTGGGAAGGTGTGGCCTGGCTCTACTGCCTTCCCTGACTTCACCAATCCTACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGA CCAGGTGCCCTTTGATGGCATGTGGATTGACATGAATGAGCCCTCTAATTTCATCAGGGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACCCTGCAGGCTGCCACCATCTGTGCTAGCTCTCACCAGTTCCTGAGCACCCACTACAATCTGCATAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCACAGGGCCCTGG TGAAGGCTAGGGGCACCAGGCCCTTTGTGATTTCTAGGAGCACTTTTGCTGGCCATGGCAGGTATGCTGGGCACTGGACTGGGGATGTGTGGTCTAGCTGGGAGCAGCTGGCTTCTTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGGGTTCCTGGGCAACACTTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTCTACC CTTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCCCAGGAGCCCTACAGCTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCACCTG TACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCTAGGCCTCTGTTCCTGGAGTTCCCCAAGGACTCTAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCCCT GCTGATCACTCCTGTGCTGCAGGCTGGGAAGGCTGAGGTGACTGGCTATTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCCCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTGGATACCATCAATGTGCACCTGAGGGCTGGCTACATCATTCCCCTGCAGGGCCCTGGCCTGACC ACTACTGAGTCTAGGCAGCAGCCCATGGCCCTGGCTGTGGCCCTGACCAAGGGGGGGGAGGCTAGGGGGGAGCTGTTTTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTACACTCAGGTGATCTTCCTGGCCAGGAACAATACCATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCC CCAGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTATAGCCCTGATACCAAGGTGCTGGATATTTGTGTGAGCCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGAC TCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCT TTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGTGGGGTGGGGGCTAGCTCTAGA

SEQ ID NO: 8: Poly-A GAA7 Nucleic Acid Sequence BGH WT

ATGGCTTTCCTGTGGCTGCTGTCTTGTTGGGCTCTGCTGGGCACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGCAGCAGCCCTGTGCTGGAGGAGACCCACCCTGCTCATCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCCCAGGCTCACCCTGGGAGACCCAGGGCTGTGCCCACTCAGTGTGATGTGCCCCCCAACAGCAGGTTTGACTGTGCTCCTGACAAGGCTATCACCCAGGAGCAGTGTGAGGCCAGGGGGTGCTGCTACATTCCTGCTAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCTCTTATCCCAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGCTACACTGCCACCCTGACCAGGACCACTCCCACCTTCTTTCCCAAGGATATTCTGACTCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCACTTCACTATCAAGGACCCTGCCAATAGGAGGTATGAGGTGCCCCTGGAGACTCCTCATGTGCATAGCAGGGCCCCTTCTCCTCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTGCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGAGCACTTCTCTGCCCAGCCAGTACATTACTGGGCTGGCTGAGCATCTGAGCCCCCTGATGCTGAGCACCTCTTGGACCAGGATCACCCTGTGGAACAGGGACCTGGCCCCCACTCCTGGGGCTAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTTCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCTCTCCAGCCCTGTCTTGGAGGAGCACTGGGGGCATTCTGGATGTGTACATTTTCCTGGGGCCTGAACCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTACCCCTTCATGCCCCCCTATTGGGGGCTGGGGTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCATTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGATAGCAGGAGGGATTTCACCTTCAACAAGGATGGCTTCAGGGACTTTCCTGCCATGGTGCAGGAGCTGCACCAGGGGGGCAGGAGGTATATGATGATTGTGGACCCTGCTATCAGCAGCTCTGGCCCTGCTGGCTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTTATCACTAATGAAACTGGCCAGCCTCTGATTGGCAAGGTCTGGCCTGGCTCTACTGCCTTCCCTGATTTTACTAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGATCAGGTGCCTTTTGATGGCATGTGGATTGATATGAATGAACCAAGCAACTTCATCAGAGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACCATTTGTGCTAGCAGCCACCAGTTCCTGAGCACCCACTACAATCTGCACAACCTGTATGGCCTGACTGAAGCCATTGCCAGCCATAGGGCCCTGGTGAAGGCCAGGGGCACTAGGCCTTTTGTGATCAGCAGGAGCACTTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCAGCTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATTCTGCAGTTTAACCTGCTGGGGGTGCCCCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTTTATCCCTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCTCAGGAGCCCTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCACCTGTATACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTCCTGGAGTTCCCCAAGGACAGCAGCACCTGGACTGTGGATCATCAGCTGCTGTGGGGGGAGGCCCTGCTGATCACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTCACTGGCTACTTCCCTCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCTCTGGGCAGCCTGCCCCCCCCCCCTGCTGCTCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCTCCCCTGGACACCATCAATGTGCACCTGAGGGCTGGCTACATTATCCCCCTGCAGGGCCCAGGGCTGACTACCACTGAGAGCAGACAGCAGCCCATGGCTCTGGCTGTGGCCCTGACCAAGGGGGGGGAAGCTAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTATACCCAGGTGATCTTCCTGGCTAGGAACAACACCATTGTCAATGAGCTGGTGAGGGTGACTTCTGAGGGGGCTGGGCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCTCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACAGCCCTGACACCAAGGTGCTGGACATCTGTGTGTCTCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGACTCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGA

SEQ ID NO: 9 : GAA8 Poly-A Nucleic Acid Sequence BGH WT

ATGGCCTTCCTGTGGCTGCTGTCTTGCTGGGCTCTGCTGGGGACCACCTTTGGCCTGCTGGTCCCCAGGGAGCTGTCTGGCTCTTCTCCTGTCCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCCACCCAGTGTGATGTGCCTCCCAACAGCAGGTTTGACTGTGCCCCTGACAAGGCCATCACCCAGGAGCAGTGTGAGGCCAGGGGCTGCTGCTATATCCCTGCCAAGCAGGGCCTGCAGGGGGCTCAGATGGGCCAGCCCTGGTGCTTCTTTCCCCCCTCTTATCCTAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGGTACACTGCCACCCTGACCAGGACCACCCCCACTTTCTTCCCTAAGGACATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAATAGGCTGCACTTTACTATCAAGGACCCTGCCAACAGGAGGTATGAGGTGCCTCTGGAGACCCCCCATGTGCATTCTAGGGCCCCCAGCCCCCTGTACTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGACAGCTGGATGGCAGGGTCCTGCTGAACACCACTGTGGCTCCCCTGTTTTTTGCTGACCAGTTCCTGCAGCTGAGCACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCACCTGAGCCCCCTGATGCTGAGCACCAGCTGGACCAGGATCACCCTGTGGAACAGGGATCTGGCTCCTACCCCTGGGGCCAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCAGCCCTGCCCTGAGCTGGAGGTCTACTGGGGGCATCCTGGATGTGTACATCTTTCTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTATCCTTTTATGCCCCCCTATTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGACAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGACAGCAGGAGGGACTTCACCTTTAACAAGGATGGCTTTAGGGACTTCCCTGCCATGGTGCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCAGCCATCAGCAGCTCTGGGCCTGCTGGGTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGGAGCACTGCCTTCCCTGATTTTACCAACCCCACTGCCCTGGCCTGGTGGGAGGATATGGTGGCTGAGTTTCATGACCAGGTGCCCTTTGATGGCATGTGGATTGACATGAATGAGCCCAGCAATTTCATCAGGGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAATCCTCCCTATGTGCCTGGGGTGGTGGGGGGCACCCTGCAGGCTGCCACCATCTGTGCCTCTAGCCACCAGTTCCTGAGCACCCACTATAACCTGCATAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGAGCCCTGGTGAAGGCCAGAGGGACCAGGCCCTTTGTGATCTCTAGGAGCACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCTCTTGGGAGCAGCTGGCCAGCTCTGTGCCAGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAATACCTCTGAAGAGCTGTGTGTGAGGTGGACTCAGCTGGGGGCCTTCTATCCCTTCATGAGGAACCACAACAGCCTGCTGTCTCTGCCCCAGGAGCCCTACAGCTTCTCTGAGCCTGCTCAGCAGGCTATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCATCTGTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTTCTGGAGTTTCCCAAGGACAGCAGCACCTGGACTGTGGACCATCAGCTGCTGTGGGGGGAGGCTCTGCTGATTACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGGTACTTCCCCCTGGGGACTTGGTATGACCTGCAGACTGTGCCTGTGGAAGCTCTGGGCAGCCTGCCCCCACCCCCTGCTGCCCCTAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGACACCATCAATGTGCACCTGAGGGCTGGCTATATCATCCCCCTGCAGGGCCCTGGGCTGACCACCACTGAGAGCAGGCAGCAGCCCATGGCCCTGGCTGTGGCCCTGACTAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGAGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAACACCATTGTGAATGAGCTGGTGAGGGTGACTTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGTCTAACTTCACCTACAGCCCTGATACTAAGGTGCTGGATATCTGTGTGAGCCTGCTGATGGGGGAGCAGTTTCTGGTGAGCTGGTGCTGACTCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGA

SEQ ID NO: 10: Nucleic acid sequence of GAA13 with poly-A BGH WT

ATGGCCTTTCTGTGGCTGCTGTCCTGCTGGGCCCTGCTGGGGACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGGAGCAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCCAGCAGGCCTGGCCCTAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCTACCCAGTGTGATGTGCCACCCAATTCTAGGTTTGACTGTGCTCCTGACAAGGCCATCACTCAGGAGCAGTGTGAAGCTAGGGGGTGCTGCTACATCCCAGCCAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTACCCTAGCTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCTACCCTGACCAGGACCACTCCTACCTTCTTCCCCAAGGACATCCTGACTCTGAGGCTGGATGTCATGATGGAGACTGAAAATAGGCTGCACTTCACCATCAAGGACCCTGCCAATAGGAGGTATGAGGTGCCTCTGGAGACCCCCCATGTGCATAGCAGGGCTCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTCATTGTGAGGAGACAGCTGGATGGGAGGGTGCTGCTGAACACTACTGTGGCTCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGTCTACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCATCTGAGCCCCCTGATGCTGAGCACCAGCTGGACCAGGATCACTCTGTGGAACAGGGATCTGGCCCCCACTCCTGGGGCCAACCTGTATGGGAGCCATCCCTTCTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCCATGGATGTGGTGCTGCAGCCTAGCCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTCTACATCTTCCTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTATCTGGATGTGGTGGGGTATCCCTTCATGCCCCCCTACTGGGGCCTGGGCTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCTCTGGATGTGCAGTGGAATGACCTGGACTATATGGATTCTAGGAGAGACTTTACTTTTAACAAGGATGGCTTCAGGGATTTCCCTGCCATGGTGCAGGAGCTGCACCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCTATTAGCAGCTCTGGCCCTGCTGGGTCTTACAGGCCTTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGCAGCACTGCCTTCCCTGACTTCACCAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGACCAGGTGCCCTTTGATGGGATGTGGATTGACATGAATGAGCCCTCTAACTTCATCAGGGGGTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACTATCTGTGCTTCTTCTCACCAGTTTCTGAGCACCCACTATAATCTGCACAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGGGCCCTGGTGAAGGCCAGGGGCACCAGGCCCTTTGTGATCAGCAGGTCTACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGTCTTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGTGGCTTTCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTTTACCCCTTCATGAGGAACCACAATAGCCTGCTGAGCCTGCCCCAGGAGCCTTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACTCTGAGGTATGCCCTGCTGCCCCATCTGTATACCCTGTTTCACCAGGCCCATGTGGCTGGGGAGACTGTGGCTAGGCCTCTGTTTCTGGAGTTCCCTAAGGACTCTAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCCCTGCTGATCACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCTCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATTCATTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGACACCATCAATGTGCACCTGAGGGCTGGGTACATCATCCCCCTGCAGGGCCCTGGCCTGACCACCACTGAGAGCAGGCAGCAGCCCATGGCCCTGGCTGTGGCTCTGACCAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGTCTCTGGAGGTGCTGGAGAGGGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAATACTATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTCCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACTCTCCTGACACCAAGGTGCTGGACATTTGTGTGTCTCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGACTCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGA

SEQ ID NO: 11: GAA2 poly-A BGH nucleic acid sequence with reduced CpG content

ATGGCTTTCCTGTGGCTGCTGAGCTGCTGGGCTCTGCTGGGCACCACCTTTGGGCTGCTGGTGCCTAGGGAGCTGTCTGGGTCTAGCCCTGTGCTGGAGGAGACTCACCCTGCCCATCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCTCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCCACCCAGTGTGATGTGCCCCCCAACAGCAGGTTTGACTGTGCCCCTGACAAGGC CATTACCCAGGAGCAGTGTGAGGCCAGGGGCTGCTGCTACATTCCAGCTAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTATCCTAGCTATAAACTG GAGAACCTGAGCAGCTCTGAGATGGGCTATACTGCCACCCTGACTAGGACTACTCCCACCTTTTTTCCTAAGGATATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGC ACTTCACTATTAAGGACCCTGCCAATAGGAGGTATGAAGTGCCTCTGGAGACTCCTCATGTGCACTCTAGGGCCCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGT GATTGTGAGGAGGCAGCTGGATGGCAGGGTGCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGAGCACCAGCCTGCCCAGCCAGTACATCACTGGGCTG GCTGAGCATCTGAGCCCTCTGATGCTGAGCACCTCTTGGACCAGGATCACCCTGTGGAATAGGGATCTGGCCCCCACCCCTGGGGCTAATCTGTATGGCTCTCATCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTTCTGCTGAACAGCAATGCCATGGATGTGGTGCTGCAGCCCTCTCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTGTACATCTTC CTGGGCCCTGAGCCCAAGTCTGTGGTCCAGCAGTATCTGGATGTGGTGGGCTACCCCTTTATGCCCCCCTATTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGGTATTCTTCTACTGCT ATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCTCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTATATGGACTCTAGGAGGGATTTCACCTTCAACAAGGATGGCTTCAG ACTTCCCTGCTATGGTCCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCCATCAGCAGCTCTGGCCCTGCTGGCAGCTATAGGCCCTATGATGAGGGCCTGAGGAGGGG GGTGTTTATCACTAATGAAACTGGGCAGCCCCTGATTGGCAAGGTGTGGCCTGGCTCTACTGCCTTCCCTGACTTCACCAACCCCACTGCTCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCAT GACCAGGTGCCTTTTGATGCATGTGGATTGACATGAATGAGCCCAGCAACTTCATCAGGGGCTCTGAGGATGGGTGCCCCAATAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGG TGGGGGGCACCCTGCAGGCTGCCACTATTTGTGCCAGCTCTCACCAGTTCCTGAGCACCCACTACAACCTGCACAATCTGTATGGCCTGACTGAGGCCATTGCCAGCCACAGGGCCCTTG AAGGCCAGGGGCACTAGGCCCTTTGTGATCTCTAGAAGCACCTTTGCTGGCCATGGGAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCCCTGGTGGGGGCTGATGTGTGGCTTCCTGGGCAACACCTCTGAAGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTCTA CCCTTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCTCAGGAGCCTTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCTCTGCTGCCCCACCTGTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTCCTGGAGTTTCCTAAGGATAGCAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGG CCCTGCTGATTACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACTTGGTATGACCTGCAGACTGTGCCTGTGGAAGCCCTGGGCAGCCTGCCTCCCCCCCCT GCTGCCCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCCCTGGACACCATTAATGTGCATCTGAGGGCTGGGTATATTATCCCCCTGCAGGGGCCTGGGCTGACT ACCACTGAGAGCAGGCAGCAGCCTATGGCCCTGGCTGTGGCTCTGACTAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGGGGGGCCTACACCCAGGTGATTTTCCTGGCCAGGAACAACACCATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAAGTGACTGTGCTGGGGGTGGCCACTGCCCCCC AGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACAGCCCTGACACCAAGGTGCTGGATATTTGTGTGAGCCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGAAG ATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTG TCCTTTCCTAATAAAATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGG

SEQ ID NO: 12: GAA5 Poly-A BGH CpG Reduced Nucleic Acid Sequence

ATGGCTTTCCTGTGGCTGCTGTCTTGCTGGGCCCTGCTGGGGACTACCTTTGGCCTGCTGGTGCCCAGGGAACTGTCTGGCTCTAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCTTCTAGGCCTGGCCCCAGGGATGCCCAGGCCCACCCTGGCAGGCCAAGGGCTGTGCCCACCCAGTGTGATGTGCCCCCCAACTCTAGATTTGATTGTGCCCCTGATAAGGCCAT CACCCAGGAGCAGTGTGAGGCTAGGGGCTGCTGCTACATCCCTGCTAAGCAGGGCCTGCAGGGGGCTCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTATCCCTCTTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCCACCCTGACCAGGACTACTCCCACCTTCTTCCCCAAGGACATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCATT TCACCATCAAGGATCCTGCCAACAGGAGGTATGAGGTGCCTCTGGAGACCCCCATGTGCACAGCAGGGCTCCTTCTCCCCTGTACTCTGTGGAGTTCTCTGAGGAACCCTTTGGGGTGA TTGTGAGGAGGCAGCTGGATGGCAGGGTCCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGATCAGTTCCTGCAGCTGTCCACTTCTCTGCCTAGCCAGTACATCACTGGGCTGGC TGAGCACCTGAGCCCTCTGATGCTGAGCACCTCTTGGACTAGGATCACCCTGTGGAACAGGGACCTGGCCCCCACCCCTGGGGCCAACCTGTATGGCAGCCACCCCTTCTATCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAATAGCAATGCTATGGATGTGGTGCTGCAGCCCAGCCCTGCCCTGTCTTGGAGGAGCACTGGGGGCATCCTGGATGTGTACATTT TCCTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTACCCCTTCATGCCTCCCTACTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGCTACAGCTCTACTGCCA TCACCAGGCAGGTGGTGGAATATGACCAGGGCCCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGACTCTAGGAGGGACTTCACCTTCAATAAGGATGGCTTCAGAGACT TCCCTGCCATGGTGCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCCATCAGCTCTTCTGGCCCTGCTGGCTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGGCAGCCCCTGATTGGGAAGGTGTGGCCTGGCTCTACTGCCTTCCCTGACTTCACCAATCCTACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCAT GACCAGGTGCCCTTTGATGGCATGTGGATTGACATGAATGAGCCCTCTAATTTCATCAGGGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGT GGGGGGCACCCTGCAGGCTGCCACCATCTGTGCTAGCTCTCACCAGTTCCTGAGCACCCACTACAATCTGCATAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCACAGGGCCCTGGTGA AGGCTAGGGGCACCAGGCCCTTTGTGATTTCTAGGAGCACTTTTGCTGGCCATGGCAGGTATGCTGGGCACTGGACTGGGGATGTGTGGTCTAGCTGGGAGCAGCTGGCTTCTTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGGGTTCCTGGGCAACACTTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTCTACC CTTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCCCAGGAGCCCTACAGCTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCACCT GTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCTAGGCCTCTGTTCCTGGAGTTCCCCAAGGACTCTAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCC CTGCTGATCACTCCTGTGCTGCAGGCTGGGAAGGCTGAGGTGACTGGCTATTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCCCCCCCCCCTG CTGCCCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTGGATACCATCAATGTGCACCTGAGGGCTGGCTACATCATTCCCCTGCAGGGCCCTGGCCTGACC ACTACTGAGTCTAGGCAGCAGCCCATGGCCCTGGCTGTGGCCCTGACCAAGGGGGGGGAGGCTAGGGGGGAGCTGTTTTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTAC ACTCAGGTGATCTTCCTGGCCAGGAACAATACCATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAG CAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTATAGCCCTGATACCAAGGTGCTGGATATTTGTGTGAGCCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGAAGATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCT TTCCTAATAAAATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGG

SEQ ID NO: 13: GAA7 poly-A BGH nucleic acid sequence with reduced CpG content

ATGGCTTTCCTGTGGCTGCTGTCTTGTTGGGCTCTGCTGGGCACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGCAGCAGCCCTGTGCTGGAGGAGACCCACCCTGCTCATCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCCCAGGCTCACCCTGGGAGACCCAGGGCTGTGCCCACTCAGTGTGATGTGCCCCCCAACAGCAGGTTTGACTGTGCTCCTGACAAGGCTATCACCCAGGAGCAGTGTGAGGCCAGGGGGTGCTGCTACATTCCTGCTAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCTCTTATCCCAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGCTACACTGCCACCCTGACCAGGACCACTCCCACCTTCTTTCCCAAGGATATTCTGACTCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCACTTCACTATCAAGGACCCTGCCAATAGGAGGTATGAGGTGCCCCTGGAGACTCCTCATGTGCATAGCAGGGCCCCTTCTCCTCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTGCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGAGCACTTCTCTGCCCAGCCAGTACATTACTGGGCTGGCTGAGCATCTGAGCCCCCTGATGCTGAGCACCTCTTGGACCAGGATCACCCTGTGGAACAGGGACCTGGCCCCCACTCCTGGGGCTAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTTCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCTCTCCAGCCCTGTCTTGGAGGAGCACTGGGGGCATTCTGGATGTGTACATTTTCCTGGGGCCTGAACCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTACCCCTTCATGCCCCCCTATTGGGGGCTGGGGTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCATTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGATAGCAGGAGGGATTTCACCTTCAACAAGGATGGCTTCAGGGACTTTCCTGCCATGGTGCAGGAGCTGCACCAGGGGGGCAGGAGGTATATGATGATTGTGGACCCTGCTATCAGCAGCTCTGGCCCTGCTGGCTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTTATCACTAATGAAACTGGCCAGCCTCTGATTGGCAAGGTCTGGCCTGGCTCTACTGCCTTCCCTGATTTTACTAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGATCAGGTGCCTTTTGATGGCATGTGGATTGATATGAATGAACCAAGCAACTTCATCAGAGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACCATTTGTGCTAGCAGCCACCAGTTCCTGAGCACCCACTACAATCTGCACAACCTGTATGGCCTGACTGAAGCCATTGCCAGCCATAGGGCCCTGGTGAAGGCCAGGGGCACTAGGCCTTTTGTGATCAGCAGGAGCACTTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCAGCTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATTCTGCAGTTTAACCTGCTGGGGGTGCCCCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTTTATCCCTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCTCAGGAGCCCTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCACCTGTATACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTCCTGGAGTTCCCCAAGGACAGCAGCACCTGGACTGTGGATCATCAGCTGCTGTGGGGGGAGGCCCTGCTGATCACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTCACTGGCTACTTCCCTCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCTCTGGGCAGCCTGCCCCCCCCCCCTGCTGCTCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCTCCCCTGGACACCATCAATGTGCACCTGAGGGCTGGCTACATTATCCCCCTGCAGGGCCCAGGGCTGACTACCACTGAGAGCAGACAGCAGCCCATGGCTCTGGCTGTGGCCCTGACCAAGGGGGGGGAAGCTAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTATACCCAGGTGATCTTCCTGGCTAGGAACAACACCATTGTCAATGAGCTGGTGAGGGTGACTTCTGAGGGGGCTGGGCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCTCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACAGCCCTGACACCAAGGTGCTGGACATCTGTGTGTCTCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGAAGATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGG

SEQ ID NO: 14: GAA8 poly-A BGH nucleic acid sequence with reduced CpG content

ATGGCCTTCCTGTGGCTGCTGTCTTGCTGGGCTCTGCTGGGGACCACCTTTGGCCTGCTGGTCCCCAGGGAGCTGTCTGGCTCTTCTCCTGTCCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCCACCCAGTGTGATGTGCCTCCCAACAGCAGGTTTGACTGTGCCCCTGACAAGGCCATCACCCAGGAGCAGTGTGAGGCCAGGGGCTGCTGCTATATCCCTGCCAAGCAGGGCCTGCAGGGGGCTCAGATGGGCCAGCCCTGGTGCTTCTTTCCCCCCTCTTATCCTAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGGTACACTGCCACCCTGACCAGGACCACCCCCACTTTCTTCCCTAAGGACATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAATAGGCTGCACTTTACTATCAAGGACCCTGCCAACAGGAGGTATGAGGTGCCTCTGGAGACCCCCCATGTGCATTCTAGGGCCCCCAGCCCCCTGTACTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGACAGCTGGATGGCAGGGTCCTGCTGAACACCACTGTGGCTCCCCTGTTTTTTGCTGACCAGTTCCTGCAGCTGAGCACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCACCTGAGCCCCCTGATGCTGAGCACCAGCTGGACCAGGATCACCCTGTGGAACAGGGATCTGGCTCCTACCCCTGGGGCCAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCAGCCCTGCCCTGAGCTGGAGGTCTACTGGGGGCATCCTGGATGTGTACATCTTTCTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTATCCTTTTATGCCCCCCTATTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGACAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGACAGCAGGAGGGACTTCACCTTTAACAAGGATGGCTTTAGGGACTTCCCTGCCATGGTGCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCAGCCATCAGCAGCTCTGGGCCTGCTGGGTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGGAGCACTGCCTTCCCTGATTTTACCAACCCCACTGCCCTGGCCTGGTGGGAGGATATGGTGGCTGAGTTTCATGACCAGGTGCCCTTTGATGGCATGTGGATTGACATGAATGAGCCCAGCAATTTCATCAGGGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAATCCTCCCTATGTGCCTGGGGTGGTGGGGGGCACCCTGCAGGCTGCCACCATCTGTGCCTCTAGCCACCAGTTCCTGAGCACCCACTATAACCTGCATAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGAGCCCTGGTGAAGGCCAGAGGGACCAGGCCCTTTGTGATCTCTAGGAGCACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCTCTTGGGAGCAGCTGGCCAGCTCTGTGCCAGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAATACCTCTGAAGAGCTGTGTGTGAGGTGGACTCAGCTGGGGGCCTTCTATCCCTTCATGAGGAACCACAACAGCCTGCTGTCTCTGCCCCAGGAGCCCTACAGCTTCTCTGAGCCTGCTCAGCAGGCTATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCATCTGTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTTCTGGAGTTTCCCAAGGACAGCAGCACCTGGACTGTGGACCATCAGCTGCTGTGGGGGGAGGCTCTGCTGATTACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGGTACTTCCCCCTGGGGACTTGGTATGACCTGCAGACTGTGCCTGTGGAAGCTCTGGGCAGCCTGCCCCCACCCCCTGCTGCCCCTAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGACACCATCAATGTGCACCTGAGGGCTGGCTATATCATCCCCCTGCAGGGCCCTGGGCTGACCACCACTGAGAGCAGGCAGCAGCCCATGGCCCTGGCTGTGGCCCTGACTAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGAGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAACACCATTGTGAATGAGCTGGTGAGGGTGACTTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGTCTAACTTCACCTACAGCCCTGATACTAAGGTGCTGGATATCTGTGTGAGCCTGCTGATGGGGGAGCAGTTTCTGGTGAGCTGGTGCTGAAGATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGG

SEQ ID NO: 15: GAA13 poly-A BGH nucleic acid sequence with reduced CpG content

ATGGCCTTTCTGTGGCTGCTGTCCTGCTGGGCCCTGCTGGGGACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGGAGCAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCCAGCAGGCCTGGCCCTAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCTACCCAGTGTGATGTGCCACCCAATTCTAGGTTTGACTGTGCTCCTGACAAGGCCATCACTCAGGAGCAGTGTGAAGCTAGGGGGTGCTGCTACATCCCAGCCAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTACCCTAGCTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCTACCCTGACCAGGACCACTCCTACCTTCTTCCCCAAGGACATCCTGACTCTGAGGCTGGATGTCATGATGGAGACTGAAAATAGGCTGCACTTCACCATCAAGGACCCTGCCAATAGGAGGTATGAGGTGCCTCTGGAGACCCCCCATGTGCATAGCAGGGCTCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTCATTGTGAGGAGACAGCTGGATGGGAGGGTGCTGCTGAACACTACTGTGGCTCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGTCTACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCATCTGAGCCCCCTGATGCTGAGCACCAGCTGGACCAGGATCACTCTGTGGAACAGGGATCTGGCCCCCACTCCTGGGGCCAACCTGTATGGGAGCCATCCCTTCTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCCATGGATGTGGTGCTGCAGCCTAGCCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTCTACATCTTCCTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTATCTGGATGTGGTGGGGTATCCCTTCATGCCCCCCTACTGGGGCCTGGGCTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCTCTGGATGTGCAGTGGAATGACCTGGACTATATGGATTCTAGGAGAGACTTTACTTTTAACAAGGATGGCTTCAGGGATTTCCCTGCCATGGTGCAGGAGCTGCACCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCTATTAGCAGCTCTGGCCCTGCTGGGTCTTACAGGCCTTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGCAGCACTGCCTTCCCTGACTTCACCAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGACCAGGTGCCCTTTGATGGGATGTGGATTGACATGAATGAGCCCTCTAACTTCATCAGGGGGTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACTATCTGTGCTTCTTCTCACCAGTTTCTGAGCACCCACTATAATCTGCACAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGGGCCCTGGTGAAGGCCAGGGGCACCAGGCCCTTTGTGATCAGCAGGTCTACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGTCTTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGTGGCTTTCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTTTACCCCTTCATGAGGAACCACAATAGCCTGCTGAGCCTGCCCCAGGAGCCTTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACTCTGAGGTATGCCCTGCTGCCCCATCTGTATACCCTGTTTCACCAGGCCCATGTGGCTGGGGAGACTGTGGCTAGGCCTCTGTTTCTGGAGTTCCCTAAGGACTCTAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCCCTGCTGATCACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCTCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATTCATTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGACACCATCAATGTGCACCTGAGGGCTGGGTACATCATCCCCCTGCAGGGCCCTGGCCTGACCACCACTGAGAGCAGGCAGCAGCCCATGGCCCTGGCTGTGGCTCTGACCAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGTCTCTGGAGGTGCTGGAGAGGGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAATACTATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTCCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACTCTCCTGACACCAAGGTGCTGGACATTTGTGTGTCTCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGAAGATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGG

>pAAV-ApoE/hAAT.fixUTR.GAA13.BGH (SEQ ID NO: 16)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCTAGTAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCT CTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATC TTGCTACCAGTGGAACAGCCACTAAGGATCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTG TGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGC CCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATACCACTTTCACAATCTGCTAGCGTTTAAACGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTTCTTTCCCCTTCTTTTCT ATGGTTAAGTTCATGTCATAGGAAGGGGAGAAGTAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATA TTTCTGCATATAAATATTTCTGCATATAAATTGTAACTGATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTCTGGTTGGGATAAGGCTGGATTATT CTGAGTCCAAGCTAGGCCCTTTTGCTAATCTTGTTCATACCTCTTATCTTCCTCCAAGCTCCTGGGCAACCTGCTGGTCTCTCTGCTGGCCCATCACTTTGGCAAAGCACGCGTGCCACCATGG CCTTTCTGTGGCTGCTGTCCTGCTGGGCCCTGCTGGGGACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGGAGCAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCCAGCAGGCCTGGCCCTAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCTACCCAGTGTGATGTGCCACCCAATTCTAGGTTTGACTGTGCTCCTGACAAGGCCATC ACTCAGGAGCAGTGTGAAGCTAGGGGGTGCTGCTACATCCCAGCCAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTACCCTAGCTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCTACCCTGACCAGGACCACTCCTACCTTCTTCCCCAAGGACATCCTGACTCTGAGGCTGGATGTCATGATGGAGACTGAAAATAGGCTGCACTTCAC CATCAAGGACCCTGCCAATAGGAGGTATGAGGTGCCTCTGGAGACCCCCATGTGCATAGCAGGGCTCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTCATTGT GAGGAGACAGCTGGATGGGAGGGTGCTGCTGAACACTACTGTGGCTCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGTCTACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAG CATCTGAGCCCCCTGATGCTGAGCACCAGCTGGACCAGGATCACTCTGTGGAACAGGGATCTGGCCCCCACTCCTGGGGCCAACCTGTATGGGAGCCATCCCTTCTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCCATGGATGTGGTGCTGCAGCCTAGCCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTCTACATCTTCCT GGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTATCTGGATGTGGTGGGGTATCCCTTCATGCCCCCCTACTGGGGCCTGGGCTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCTCTGGATGTGCAGTGGAATGACCTGGACTATATGGATTCTAGGAGAGACTTTACTTTTAACAAGGATGGCTTCAGGGAT TTCCCTGCCATGGTGCAGGAGCTGCACCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCTATTAGCAGCTCTGGCCCTGCTGGGTCTTACAGGCCTTATGATGAGGGCCTGAGGAGGGGG GTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGCAGCACTGCCTTCCCTGACTTCACCAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCAT GACCAGGTGCCCTTTGATGGGATGTGGATTGACATGAATGAGCCCTCTAACTTCATCAGGGGGTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACTATCTGTGCTTCTTCTCACCAGTTTCTGAGCACCCACTATAATCTGCACAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGGGCCCTGG TGAAGGCCAGGGGCACCAGGCCCTTTGTGATCAGCAGGTCTACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGTCTTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGGCTTTCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCT TTTACCCCTTCATGAGGAACCACAATAGCCTGCTGAGCCTGCCCCAGGAGCCTTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACTCTGAGGTATGCCCTGCTGCC CCATCTGTATACCCTGTTTCACCAGGCCCATGTGGCTGGGGAGACTGTGGCTAGGCCTCTGTTTCTGGAGTTCCCTAAGGACTCTAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGG GAGGCCCTGCTGATCACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCTCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATTCATTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGACACCATCAATGTGCACCTGAGGGCTGGGTACATCATCCCCCTGCAGGGCCCT GGCCTGACCACCACTGAGAGCAGGCAGCAGCCCATGGCCCTGGCTGTGGCTCTGACCAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGTCTCTGGAGGTGCTGGAGAGGGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAATACTATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGTGG CCACTGCCCCCCAGCAGGTCCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACTCTCCTGACACCAAGGTGCTGGACATTTGTGTGTCTCTGCTGATGGGGGAGCAGTTCCCTGGTGAG CTGGTGCTGAAGATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCTGACCCTGGAAGGTG CCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGGCTTCTGAGGCAGAAAGAACCAGCTGGGGCTCGAGATCCACTAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGC GCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

>pAAV-ApoE/hAAT.GAA13.BGH (SEQ ID NO: 17)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCTAGTAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCT CTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATC TTGCTACCAGTGGAACAGCCACTAAGGATCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTG TGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGC CCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGG GAGAAGTAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATT GTAACTGATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTCTGGTTGGGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCTTGTTCATACCTCTTATCTTCCTCCCACAGCTCCTGGGCAACCTGCTGGTCTCTCTGCTGGCCCATCACTTTGGCAAAGCACGCGTGCCACCATGGCCTTTCTGTGGCTGCTGTCCTGCTGGG CCCTGCTGGGGACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGGAGCAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCCAGCAGGCCTGGCCCTAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCTACCCAGTGTGATGTGCCACCCAATTCTAGGTTTGACTGTGCTCCTGACAAGGCCATCACTCAGGAGCAGTGTGAAGCTAGGGGG TGCTGCTACATCCCAGCCAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTACCCTAGCTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCTACCCTGACCAGGACCACTCCTACCTTCTTCCCCAAGGACATCCTGACTCTGAGGCTGGATGTCATGATGGAGACTGAAAATAGGCTGCACTTCACCATCAAGGACCCTGCCAATAGGAGGTA TGAGGTGCCTCTGGAGACCCCCATGTGCATAGCAGGGCTCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTCATTGTGAGGACAGCTGGATGGGAGGGTGCTG CTGAACACTACTGTGGCTCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGTCTACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCATCTGAGCCCCCTGATGCTGAGCACCAG CTGGACCAGGATCACTCTGTGGAACAGGGATCTGGCCCCCACTCCTGGGGCCAACCTGTATGGGAGCCATCCCTTCTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCT GAACAGCAATGCCATGGATGTGGTGCTGCAGCCTAGCCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTCTACATCTTCCTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTA TCTGGATGTGGTGGGGTATCCCTTCATGCCCCCCTACTGGGGCCTGGGCTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCTCTGGATGTGCAGTGGAATGACCTGGACTATATGGATTCTAGGAGAGACTTTACTTTTAACAAGGATGGCTTCAGGGATTTCCCTGCCATGGTGCAGGAGCTGCACCAGGGGG GCAGGAGGTACATGATGATTGTGGACCCTGCTATTAGCAGCTCTGGCCCTGCTGGGTCTTACAGGCCTTATGATGAGGGCCTGAGGAGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCT GATTGGCAAGGTGTGGCCTGGCAGCACTGCCTTCCCTGACTTCACCAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGACCAGGTGCCCTTTGATGGGATGTGGATT GACATGAATGAGCCCTCTAACTTCATCAGGGGGTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACTATCTGTGCTTCTTCTCACCAGTTTCTGAGCACCCACTATAATCTGCACAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGGGGCCCTGGTGAAGGCCAGGGGCACCAGGCCCTTTGTGATC AGCAGGTCTACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGTCTTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGGCTTTCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTTTACCCCTTCATGAGGAACCACAATAGCCTGCT A GCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCTCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATTCATTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGACACCATCAATGTGCACCTGAGGGCTGGGTACATCATCCCCCTGCAGGGCCCTGGCCTGACCACCACTGAGAGCAGGCAGCAGCCCAT GGCCCTGGCTGTGGCTCTGACCAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGTCTCTGGAGGTGCTGGAGAGGGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAATACTATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTCCTGAGCAATGGGG TGCCTGTGAGCAACTTCACCTACTCTCCTGACACCAAGGTGCTGGACATTTGTGTCTCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGAAGATCTAGAGCTGAATTCCTGCA GCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGG AAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGGCTTCTGAGGCAGAAAGAACCAGCTGGGGCTCGAGATCCACTAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGG TCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

>pAAV-ApoE/hAAT.GAA7.BGH (SEQ ID NO: 18)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCTAGTAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATCTTGCTACCAGTGGAACAGCCACTAAGGATTCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAGTAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATTGTAACTGATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTCTGGTTGGGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCTTGTTCATACCTCTTATCTTCCTCCCACAGCTCCTGGGCAACCTGCTGGTCTCTCTGCTGGCCCATCACTTTGGCAAAGCACGCGTGCCACCATGGCTTTCCTGTGGCTGCTGTCTTGTTGGGCTCTGCTGGGCACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGCAGCAGCCCTGTGCTGGAGGAGACCCACCCTGCTCATCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCCCAGGCTCACCCTGGGAGACCCAGGGCTGTGCCCACTCAGTGTGATGTGCCCCCCAACAGCAGGTTTGACTGTGCTCCTGACAAGGCTATCACCCAGGAGCAGTGTGAGGCCAGGGGGTGCTGCTACATTCCTGCTAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCTCTTATCCCAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGCTACACTGCCACCCTGACCAGGACCACTCCCACCTTCTTTCCCAAGGATATTCTGACTCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCACTTCACTATCAAGGACCCTGCCAATAGGAGGTATGAGGTGCCCCTGGAGACTCCTCATGTGCATAGCAGGGCCCCTTCTCCTCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTGCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGAGCACTTCTCTGCCCAGCCAGTACATTACTGGGCTGGCTGAGCATCTGAGCCCCCTGATGCTGAGCACCTCTTGGACCAGGATCACCCTGTGGAACAGGGACCTGGCCCCCACTCCTGGGGCTAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTTCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCTCTCCAGCCCTGTCTTGGAGGAGCACTGGGGGCATTCTGGATGTGTACATTTTCCTGGGGCCTGAACCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTACCCCTTCATGCCCCCCTATTGGGGGCTGGGGTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCATTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGATAGCAGGAGGGATTTCACCTTCAACAAGGATGGCTTCAGGGACTTTCCTGCCATGGTGCAGGAGCTGCACCAGGGGGGCAGGAGGTATATGATGATTGTGGACCCTGCTATCAGCAGCTCTGGCCCTGCTGGCTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTTATCACTAATGAAACTGGCCAGCCTCTGATTGGCAAGGTCTGGCCTGGCTCTACTGCCTTCCCTGATTTTACTAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGATCAGGTGCCTTTTGATGGCATGTGGATTGATATGAATGAACCAAGCAACTTCATCAGAGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACCATTTGTGCTAGCAGCCACCAGTTCCTGAGCACCCACTACAATCTGCACAACCTGTATGGCCTGACTGAAGCCATTGCCAGCCATAGGGCCCTGGTGAAGGCCAGGGGCACTAGGCCTTTTGTGATCAGCAGGAGCACTTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCAGCTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATTCTGCAGTTTAACCTGCTGGGGGTGCCCCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTTTATCCCTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCTCAGGAGCCCTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCACCTGTATACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTCCTGGAGTTCCCCAAGGACAGCAGCACCTGGACTGTGGATCATCAGCTGCTGTGGGGGGAGGCCCTGCTGATCACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTCACTGGCTACTTCCCTCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCTCTGGGCAGCCTGCCCCCCCCCCCTGCTGCTCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCTCCCCTGGACACCATCAATGTGCACCTGAGGGCTGGCTACATTATCCCCCTGCAGGGCCCAGGGCTGACTACCACTGAGAGCAGACAGCAGCCCATGGCTCTGGCTGTGGCCCTGACCAAGGGGGGGGAAGCTAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTATACCCAGGTGATCTTCCTGGCTAGGAACAACACCATTGTCAATGAGCTGGTGAGGGTGACTTCTGAGGGGGCTGGGCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCTCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACAGCCCTGACACCAAGGTGCTGGACATCTGTGTGTCTCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGAAGATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGGCTTCTGAGGCAGAAAGAACCAGCTGGGGCTCGAGATCCACTAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

>pAAV-ApoE/hAAT.GAA8.BGH (SEQ ID NO: 19)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCTAGTAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCT CTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATC TTGCTACCAGTGGAACAGCCACTAAGGATCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTG TGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGC CCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGG GAGAAGTAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATT GTAACTGATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTCTGGTTGGGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCTTGTTCATACCTCTTATCTTCCTCCCACAGCTCCTGGGCAACCTGCTGGTCTCTCTGCTGGCCCATCACTTTGGCAAAGCACGCGTGCCACCATGGCCTTCCTGTGGCTGCTGTCTTGCTGGG CTCTGCTGGGGACCACCTTTGGCCTGCTGGTCCCCAGGGAGCTGTCTGGCTCTTCTCCTGTCCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCCACCCAGTGTGATGTGCCTCCCCAACAGCAGGTTTGACTGTGCCCCTGACAAGGCCATCACCCAGGAGCAGTGTGAGGCCAGGGGCT GCTGCTATATCCCTGCCAAGCAGGGCCTGCAGGGGGCTCAGATGGGCCAGCCCTGGTGCTTCTTTCCCCCCTTATCCTAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGGTACACTGCCACCCTGACCAGGACCACCCCCACTTTCTTCCCTAAGGACATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAATAGGCTGCACTTTACTATCAAGGACCCTGCCAACAGGAGG TATGAGGTGCCTCTGGAGACCCCCATGTGCATTCTAGGGCCCCAGCCCCCTGTACTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGACAGCTGGATGGCAGGGTC CTGCTGAACACCACTGTGGCTCCCCTGTTTTTTGCTGACCAGTTCCTGCAGCTGAGCACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCACCTGAGCCCCCTGATGCTGAGCA CCAGCTGGACCAGGATCACCCTGTGGAACAGGGATCTGGCTCCTACCCCTGGGGCCAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCAGCCCTGCCCTGAGCTGGAGGTCTACTGGGGGCATCCTGGATGTGTACATCTTTCTGGGGCCTGAGCCCAAGTCTGTGGTGCA GCAGTACCTGGATGTGGTGGGCTATCCTTTTATGCCCCCCTATTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGACAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGACAGCAGGAGGGACTTCACCTTTAACAAGGATGGCTTTAGGGACTTCCCTGCCATGGTGCAGGAGCTGCAT CAGGGGGGCAGGAGGTACATGATGATTGTGGACCCAGCCATCAGCAGCTCTGGGCCTGCTGGGTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGGAGCACTGCCTTCCCTGATTTTACCAACCCCACTGCCCTGGCCTGGTGGGAGGATATGGTGGCTGAGTTTCATGACCAGGTGCCCTTTGATGGCAT GTGGATTGACATGAATGAGCCCAGCAATTTCATCAGGGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAATCCTCCCTATGTGCCTGGGGTGGTGGGGGGCACCCTGCAGGCTGCCACCATCTGTGCCTCTAGCCACCAGTTCCTGAGCACCCACTATAACCTGCATAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGAGCCCTGGTGAAGGCCAGAGGGACCAGGCCC TTTGTGATCTCTAGGAGCACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCTCTTGGGAGCAGCTGGCCAGCTCTGTGCCAGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGGCTTCCTGGGCAATACCTCTGAAGAGCTGTGTGTGAGGTGGACTCAGCTGGGGGCCTTCTATCCCTTCATGAGGAACCACAACAGC CTGCTGTCTCTGCCCCAGGAGCCCTACAGCTTCTCTGAGCCTGCTCAGCAGGCTATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCATCTGTACACCCTGTTCCACCAGGCC CATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTTCTGGAGTTTCCCAAGGACAGCAGCACCTGGACTGTGGACCATCAGCTGCTGTGGGGGGAGGCTCTGCTGATTACCCCTGTGCTGC AGGCTGGCAAGGCTGAGGTGACTGGGTACTTCCCCCTGGGGACTTGGTATGACCTGCAGACTGTGCCTGTGGAAGCTCTGGGCAGCCTGCCCCCACCCCCTGCTGCCCCTAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTGGACACCATCAATGTGCACCTGAGGGCTGGCTATATCATCCCCCTGCAGGGCCCTGGGCTGACCACCACTGAGAGCAGGCA GCAGCCCATGGCCCTGGCTGTGGCCCTGACTAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGAGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAACACCATTGTGAATGAGCTGGTGAGGGTGACTTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTGCTGAG CAATGGGGTGCCTGTGTCTAACTTCACCTACAGCCCTGATACTAAGGTGCTGGATATCTGTGTGAGCCTGCTGATGGGGGAGCAGTTTCTGGTGAGCTGGTGCTGAAGATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAA ATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGGCTTCTGAGGCAGAAAGAACCAGCTGGGGCTCGAGATCCACTAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGA CCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

>pAAV-ApoE/hAAT.GAA2.wtBGH (SEQ ID NO: 20)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATCTTGCTACCAGTGGAACAGCCACTAAGGATTCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAGTAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATTGTAACTGATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTCTGGTTGGGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCTTGTTCATACCTCTTATCTTCCTCCCACAGCTCCTGGGCAACCTGCTGGTCTCTCTGCTGGCCCATCACTTTGGCAAAGCACGCGTGCCACCATGGCTTTCCTGTGGCTGCTGAGCTGCTGGGCTCTGCTGGGCACCACCTTTGGGCTGCTGGTGCCTAGGGAGCTGTCTGGGTCTAGCCCTGTGCTGGAGGAGACTCACCCTGCCCATCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCTCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCCACCCAGTGTGATGTGCCCCCCAACAGCAGGTTTGACTGTGCCCCTGACAAGGCCATTACCCAGGAGCAGTGTGAGGCCAGGGGCTGCTGCTACATTCCAGCTAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTATCCTAGCTATAAACTGGAGAACCTGAGCAGCTCTGAGATGGGCTATACTGCCACCCTGACTAGGACTACTCCCACCTTTTTTCCTAAGGATATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCACTTCACTATTAAGGACCCTGCCAATAGGAGGTATGAAGTGCCTCTGGAGACTCCTCATGTGCACTCTAGGGCCCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTGCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGAGCACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCATCTGAGCCCTCTGATGCTGAGCACCTCTTGGACCAGGATCACCCTGTGGAATAGGGATCTGGCCCCCACCCCTGGGGCTAATCTGTATGGCTCTCATCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTTCTGCTGAACAGCAATGCCATGGATGTGGTGCTGCAGCCCTCTCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTGTACATCTTCCTGGGCCCTGAGCCCAAGTCTGTGGTCCAGCAGTATCTGGATGTGGTGGGCTACCCCTTTATGCCCCCCTATTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGGTATTCTTCTACTGCTATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCTCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTATATGGACTCTAGGAGGGATTTCACCTTCAACAAGGATGGCTTCAGGGACTTCCCTGCTATGGTCCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCCATCAGCAGCTCTGGCCCTGCTGGCAGCTATAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTTATCACTAATGAAACTGGGCAGCCCCTGATTGGCAAGGTGTGGCCTGGCTCTACTGCCTTCCCTGACTTCACCAACCCCACTGCTCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGACCAGGTGCCTTTTGATGGCATGTGGATTGACATGAATGAGCCCAGCAACTTCATCAGGGGCTCTGAGGATGGGTGCCCCAATAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACCCTGCAGGCTGCCACTATTTGTGCCAGCTCTCACCAGTTCCTGAGCACCCACTACAACCTGCACAATCTGTATGGCCTGACTGAGGCCATTGCCAGCCACAGGGCCCTGGTGAAGGCCAGGGGCACTAGGCCCTTTGTGATCTCTAGAAGCACCTTTGCTGGCCATGGGAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCCCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAACACCTCTGAAGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTCTACCCTTTCATGAGGAACCACAACAGCCTGCTGAGCCTGCCTCAGGAGCCTTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCTCTGCTGCCCCACCTGTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTCCTGGAGTTTCCTAAGGATAGCAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCCCTGCTGATTACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACTTGGTATGACCTGCAGACTGTGCCTGTGGAAGCCCTGGGCAGCCTGCCTCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCCCTGGACACCATTAATGTGCATCTGAGGGCTGGGTATATTATCCCCCTGCAGGGGCCTGGGCTGACTACCACTGAGAGCAGGCAGCAGCCTATGGCCCTGGCTGTGGCTCTGACTAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTACACCCAGGTGATTTTCCTGGCCAGGAACAACACCATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAAGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACAGCCCTGACACCAAGGTGCTGGATATTTGTGTGAGCCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGACTCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGACTCGAGATCCACTAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

>pAAV-ApoE/hAAT.GAA5.wtBGH (SEQ ID NO: 21)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCT GAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATCTTGC TACCAGTGGAACAGCCACTAAGGATCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCC CCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAG TAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATTGTAACT GATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTCTGGTTGGGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCTTGTTCATACCTCTTATCTTCCTCCCACAGCTCCTGGGCAACCTGCTGGTCTCTCTGCTGGCCCATCACTTTGGCAAAGCACGCGTGCCACCATGGCTTTCCTGTGGCTGCTGTCTTGCTGGGCC CTGCTGGGGACTACCTTTGGCCTGCTGGTGCCCAGGGAACTGTCTGGCTCTAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCTTCTAGGCCTGGCCCCAGGGATGCCCA GGCCCACCCTGGCAGGCCAAGGGCTGTGCCCACCCAGTGTGATGTGCCCCCCAACTCTAGATTTGATTGTGCCCCTGATAAGGCCATCACCCAGGAGCAGTGTGAGGCTAGGGGCTGCTGCTA CATCCCTGCTAAGCAGGGCCTGCAGGGGGCTCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTATCCCTCTTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCCACCCTGACCAGGACTACTCCCACCTTCTTCCCCAAGGACATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCATTTCACCATCAAGGATCCTGCCAACAGGAGGTATGA GGTGCCTCTGGAGACCCCCCATGTGCACAGCAGGGCTCCTTCTCCCCTGTACTCTGTGGAGTTCTCTGAGGAACCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTCCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGATCAGTTCCTGCAGCTGTCCACTTCTCTGCCTAGCCAGTACATCACTGGGCTGGCTGAGCACCTGAGCCCTCTGATGCTGAGCACC TCTTGGACTAGGATCACCCTGTGGAACAGGGACCTGGCCCCCACCCCTGGGGCCAACCTGTATGGCAGCCACCCCTTCTATCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTG CTGAATAGCAATGCTATGGATGTGGTGCTGCAGCCCAGCCCTGCCCTGTCTTGGAGGAGCACTGGGGGCATCCTGGATGTGTACATTTTCCTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAG TACCTGGATGTGGTGGGCTACCCCTTCATGCCTCCCTACTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGCTACAGCTCTACTGCCATCACCAGGCAGGTGGTGGAATATGACCAGGGCCCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGACTCTAGGAGGGACTTCACCTTCAATAAGGATGGCTTCAGAGACTTCCCTGCCATGGTGCAGGAGCTGCATCAGGGGGG CAGGAGGTACATGATGATTGTGGACCCTGCCATCAGCTCTTCTGGCCCTGCTGGCTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGGCAGCCCCTGATTGGGAAGGTGTGGCCTGGCTCTACTGCCTTCCCTGACTTCACCAATCCTACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGACCAGGTGCCCTTTGATGGCATGTGGATT GACATGAATGAGCCCTCTAATTTCATCAGGGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACCCTGCAGGCTGCCACCATCTGTGCTAGCTCTCACCAGTTCCTGAGCACCCACTACAATCTGCATAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCACAGGGCCCTGGTGAAGGCTAGGGGCACCAGGCCCTTTGTG ATTTCTAGGAGCACTTTTGCTGGCCATGGCAGGTATGCTGGGCACTGGACTGGGGATGTGTGGTCTAGCTGGGAGCAGCTGGCTTCTTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGGGTTCCTGGGCAACACTTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTCTACCCTTTCATGAGGAACCACAACAGCCTGCTGAG CCTGCCCCAGGAGCCCTACAGCTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCACCTGTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCTAGGCCTCTGTTCCTGGAGTTCCCCAAGGACTCTAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCCCTGCTGATCACTCCTGTGCTGCAGGCTGGG AAGGCTGAGGTGACTGGCTATTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCCCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGATACCATCAATGTGCACCTGAGGGCTGGCTACATCATTCCCCTGCAGGGCCCTGGCCTGACCACTACTGAGTCTAGGCAGCAGCCCATGG CCCTGGCTGTGGCCCTGACCAAGGGGGGGGAGGCTAGGGGGGAGCTGTTTTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTACACTCAGGTGATCTTCCTGGCCAGGAAC AATACCATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTG AGCAACTTCACCTATAGCCCTGATACCAAGGTGCTGGATATTTGTGTGAGCCTGCTGATGGGGGAGCAGTTCCCTGGTGAGCTGGTGCTGACTCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCG CATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGACTCGAGATCCACTAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

>pAAV-ApoE/hAAT.GAA7.wtBGH (SEQ ID NO: 22)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCT GAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATCTTGC TACCAGTGGAACAGCCACTAAGGATCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCC CCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAG TAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATTGTAACT GATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTCTGGTTGGGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCTTGTTCATACCTCTTATCTTCCTCCCACAGCTCCTGGGCAACCTGCTGGTCTCTCTGCTGGCCCATCACTTTGGCAAAGCACGCGTGCCACCATGGCTTTCCTGTGGCTGCTGTCTTGTTGGGC TCTGCTGGGCACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGCAGCAGCCCTGTGCTGGAGGAGACCCACCCTGCTCATCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCC CAGGCTCACCCTGGGAGACCCAGGGCTGTGCCCACTCAGTGTGATGTGCCCCCCAACAGCAGGTTTGACTGTGCTCCTGACAAGGCTATCACCCAGGAGCAGTGTGAGGCCAGGGGGTGCT GCTACATTCCTGCTAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCTCTTATCCCAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGCTACACTGCCACCCTGACCAGGACCACTCCCACCTTCTTTCCCAAGGATATTCTGACTCTGAGGCTGGATGTGATGATGGAGACTGAGAACAGGCTGCACTTCACTATCAAGGACCCTGCCAATAGGAGGTA TGAGGTGCCCCTGGAGACTCCTCATGTGCATAGCAGGGCCCCTTCTCCTCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGGCAGCTGGATGGCAGGGTGCTGCTGAACACCACTGTGGCCCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGAGCACTTCTCTGCCCAGCCAGTACATTACTGGGCTGGCTGAGCATCTGAGCCCCCTGATGCTGA GCACCTCTTGGACCAGGATCACCCTGTGGAACAGGGACCTGGCCCCCACTCCTGGGGCTAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTG TTTCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCTCTCCAGCCCTGTCTTGGAGGAGCACTGGGGGCATTCTGGATGTGTACATTTTCCTGGGGCCTGAACCCAAGTCTGTGGTG CAGCAGTACCTGGATGTGGTGGGCTACCCCTTCATGCCCCCCTATTGGGGGCTGGGGTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCATTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGATAGCAGGAGGGATTTCACCTTCAACAAGGATGGCTTCAGGGACTTTCCTGCCATGGTGCAGGAGCTG CACCAGGGGGGCAGGAGGTATATGATGATTGTGGACCCTGCTATCAGCAGCTCTGGCCCTGCTGGCTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTTATCACTAATGAAACTGGCCAGCCTCTGATTGGCAAGGTCTGGCCTGGCTCTACTGCCTTCCCTGATTTTACTAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGATCAGGTGCCTTTTG ATGGCATGTGGATTGATATGAATGAACCAAGCAACTTCATCAGAGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACCATTTGTGCTAGCAGCCACCAGTTCCTGAGCACCCACTACAATCTGCACAACCTGTATGGCCTGACTGAAGCCATTGCCAGCCATAGGGGCCCTGGTGAAGGCCAGGGGCACTAG GCCTTTTGTGATCAGCAGGAGCACTTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCAGCTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATTCTGCAGTTTAACCTGCTGGGGGTGCCCCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTTTATCCCTTCATGAGGAAC CACAACAGCCTGCTGAGCCTGCCTCAGGAGCCCTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCACCTGTATACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTCCTGGAGTTCCCCAAGGACAGCAGCACCTGGACTGTGGATCATCAGCTGCTGTGGGGGGAGGCCCTGCTGATCACCCC TGTGCTGCAGGCTGGCAAGGCTGAGGTCACTGGCTACTTCCCTCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCTCTGGGCAGCCTGCCCCCCCCCTGCTGCTCCCAGGGA GCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCTCCCCTGGACACCATCAATGTGCACCTGAGGGCTGGCTACATTATCCCCCTGCAGGGCCCAGGGCTGACTACCACTGAGAGC AGACAGCAGCCCATGGCTCTGGCTGTGGCCCTGACCAAGGGGGGGGAAGCTAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGGGGGGCCTATACCCAGGTGAT CTTCCTGGCTAGGAACAACACCATTGTCAATGAGCTGGTGAGGGTGACTTCTGAGGGCTGGGCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCTCCCCAGCAGGTGCTGAG CAATGGGGTGCCTGTGAGCAACTTCACCTACAGCCCTGACACCAAGGTGCTGGACATCTGTGTGTCTCTGCTGATGGGGGGGAGCAGTTCCCTGGTGAGCTGGTGCTGACTCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTA ATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGACTCGAGATCCACTAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

>pAAV-ApoE/hAAT.GAA8.wtBGH (SEQ ID NO: 23)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATCTTGCTACCAGTGGAACAGCCACTAAGGATTCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAGTAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATTGTAACTGATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTCTGGTTGGGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCTTGTTCATACCTCTTATCTTCCTCCCACAGCTCCTGGGCAACCTGCTGGTCTCTCTGCTGGCCCATCACTTTGGCAAAGCACGCGTGCCACCATGGCCTTCCTGTGGCTGCTGTCTTGCTGGGCTCTGCTGGGGACCACCTTTGGCCTGCTGGTCCCCAGGGAGCTGTCTGGCTCTTCTCCTGTCCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCTAGCAGGCCTGGCCCCAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCCACCCAGTGTGATGTGCCTCCCAACAGCAGGTTTGACTGTGCCCCTGACAAGGCCATCACCCAGGAGCAGTGTGAGGCCAGGGGCTGCTGCTATATCCCTGCCAAGCAGGGCCTGCAGGGGGCTCAGATGGGCCAGCCCTGGTGCTTCTTTCCCCCCTCTTATCCTAGCTATAAGCTGGAGAACCTGAGCAGCTCTGAGATGGGGTACACTGCCACCCTGACCAGGACCACCCCCACTTTCTTCCCTAAGGACATCCTGACCCTGAGGCTGGATGTGATGATGGAGACTGAGAATAGGCTGCACTTTACTATCAAGGACCCTGCCAACAGGAGGTATGAGGTGCCTCTGGAGACCCCCCATGTGCATTCTAGGGCCCCCAGCCCCCTGTACTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTGATTGTGAGGAGACAGCTGGATGGCAGGGTCCTGCTGAACACCACTGTGGCTCCCCTGTTTTTTGCTGACCAGTTCCTGCAGCTGAGCACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCACCTGAGCCCCCTGATGCTGAGCACCAGCTGGACCAGGATCACCCTGTGGAACAGGGATCTGGCTCCTACCCCTGGGGCCAACCTGTATGGCTCTCACCCCTTTTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCTATGGATGTGGTGCTGCAGCCCAGCCCTGCCCTGAGCTGGAGGTCTACTGGGGGCATCCTGGATGTGTACATCTTTCTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTACCTGGATGTGGTGGGCTATCCTTTTATGCCCCCCTATTGGGGCCTGGGCTTCCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGACAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCCCTGGATGTGCAGTGGAATGACCTGGACTACATGGACAGCAGGAGGGACTTCACCTTTAACAAGGATGGCTTTAGGGACTTCCCTGCCATGGTGCAGGAGCTGCATCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCAGCCATCAGCAGCTCTGGGCCTGCTGGGTCTTACAGGCCCTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGGAGCACTGCCTTCCCTGATTTTACCAACCCCACTGCCCTGGCCTGGTGGGAGGATATGGTGGCTGAGTTTCATGACCAGGTGCCCTTTGATGGCATGTGGATTGACATGAATGAGCCCAGCAATTTCATCAGGGGCTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAATCCTCCCTATGTGCCTGGGGTGGTGGGGGGCACCCTGCAGGCTGCCACCATCTGTGCCTCTAGCCACCAGTTCCTGAGCACCCACTATAACCTGCATAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGAGCCCTGGTGAAGGCCAGAGGGACCAGGCCCTTTGTGATCTCTAGGAGCACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGAGCTCTTGGGAGCAGCTGGCCAGCTCTGTGCCAGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGTGGCTTCCTGGGCAATACCTCTGAAGAGCTGTGTGTGAGGTGGACTCAGCTGGGGGCCTTCTATCCCTTCATGAGGAACCACAACAGCCTGCTGTCTCTGCCCCAGGAGCCCTACAGCTTCTCTGAGCCTGCTCAGCAGGCTATGAGGAAGGCCCTGACCCTGAGGTATGCCCTGCTGCCCCATCTGTACACCCTGTTCCACCAGGCCCATGTGGCTGGGGAGACTGTGGCCAGGCCCCTGTTTCTGGAGTTTCCCAAGGACAGCAGCACCTGGACTGTGGACCATCAGCTGCTGTGGGGGGAGGCTCTGCTGATTACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGGTACTTCCCCCTGGGGACTTGGTATGACCTGCAGACTGTGCCTGTGGAAGCTCTGGGCAGCCTGCCCCCACCCCCTGCTGCCCCTAGGGAGCCTGCCATCCACTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGACACCATCAATGTGCACCTGAGGGCTGGCTATATCATCCCCCTGCAGGGCCCTGGGCTGACCACCACTGAGAGCAGGCAGCAGCCCATGGCCCTGGCTGTGGCCCTGACTAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGAGCCTGGAGGTGCTGGAGAGAGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAACACCATTGTGAATGAGCTGGTGAGGGTGACTTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTGCTGAGCAATGGGGTGCCTGTGTCTAACTTCACCTACAGCCCTGATACTAAGGTGCTGGATATCTGTGTGAGCCTGCTGATGGGGGAGCAGTTTCTGGTGAGCTGGTGCTGACTCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGACTCGAGATCCACTAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

>pAAV-ApoE/hAAT.GAA13.wtBGH (SEQ ID NO: 24)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATCTTGCTACCAGTGGAACAGCCACTAAGGATTCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAGTAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCATTTGTAATTTTAAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTGCACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATTGTAACTGATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTCTGGTTGGGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCTTGTTCATACCTCTTATCTTCCTCCCACAGCTCCTGGGCAACCTGCTGGTCTCTCTGCTGGCCCATCACTTTGGCAAAGCACGCGTGCCACCATGGCCTTTCTGTGGCTGCTGTCCTGCTGGGCCCTGCTGGGGACCACCTTTGGCCTGCTGGTGCCCAGGGAGCTGTCTGGGAGCAGCCCAGTGCTGGAGGAGACCCACCCTGCCCACCAGCAGGGGGCCAGCAGGCCTGGCCCTAGGGATGCCCAGGCCCACCCTGGCAGGCCCAGGGCTGTGCCTACCCAGTGTGATGTGCCACCCAATTCTAGGTTTGACTGTGCTCCTGACAAGGCCATCACTCAGGAGCAGTGTGAAGCTAGGGGGTGCTGCTACATCCCAGCCAAGCAGGGCCTGCAGGGGGCCCAGATGGGCCAGCCCTGGTGCTTCTTCCCCCCCAGCTACCCTAGCTACAAGCTGGAGAATCTGAGCAGCTCTGAGATGGGCTACACTGCTACCCTGACCAGGACCACTCCTACCTTCTTCCCCAAGGACATCCTGACTCTGAGGCTGGATGTCATGATGGAGACTGAAAATAGGCTGCACTTCACCATCAAGGACCCTGCCAATAGGAGGTATGAGGTGCCTCTGGAGACCCCCCATGTGCATAGCAGGGCTCCCAGCCCCCTGTATTCTGTGGAGTTCTCTGAGGAGCCCTTTGGGGTCATTGTGAGGAGACAGCTGGATGGGAGGGTGCTGCTGAACACTACTGTGGCTCCCCTGTTCTTTGCTGACCAGTTCCTGCAGCTGTCTACCAGCCTGCCCAGCCAGTACATCACTGGGCTGGCTGAGCATCTGAGCCCCCTGATGCTGAGCACCAGCTGGACCAGGATCACTCTGTGGAACAGGGATCTGGCCCCCACTCCTGGGGCCAACCTGTATGGGAGCCATCCCTTCTACCTGGCCCTGGAGGATGGGGGCTCTGCCCATGGGGTGTTCCTGCTGAACAGCAATGCCATGGATGTGGTGCTGCAGCCTAGCCCTGCCCTGAGCTGGAGGAGCACTGGGGGCATCCTGGATGTCTACATCTTCCTGGGGCCTGAGCCCAAGTCTGTGGTGCAGCAGTATCTGGATGTGGTGGGGTATCCCTTCATGCCCCCCTACTGGGGCCTGGGCTTTCACCTGTGCAGGTGGGGCTACAGCAGCACTGCCATCACCAGGCAGGTGGTGGAGAACATGACCAGGGCCCACTTCCCTCTGGATGTGCAGTGGAATGACCTGGACTATATGGATTCTAGGAGAGACTTTACTTTTAACAAGGATGGCTTCAGGGATTTCCCTGCCATGGTGCAGGAGCTGCACCAGGGGGGCAGGAGGTACATGATGATTGTGGACCCTGCTATTAGCAGCTCTGGCCCTGCTGGGTCTTACAGGCCTTATGATGAGGGCCTGAGGAGGGGGGTGTTCATCACCAATGAGACTGGCCAGCCCCTGATTGGCAAGGTGTGGCCTGGCAGCACTGCCTTCCCTGACTTCACCAACCCCACTGCCCTGGCCTGGTGGGAGGACATGGTGGCTGAGTTCCATGACCAGGTGCCCTTTGATGGGATGTGGATTGACATGAATGAGCCCTCTAACTTCATCAGGGGGTCTGAGGATGGCTGCCCCAACAATGAGCTGGAGAACCCCCCCTATGTGCCTGGGGTGGTGGGGGGCACTCTGCAGGCTGCCACTATCTGTGCTTCTTCTCACCAGTTTCTGAGCACCCACTATAATCTGCACAACCTGTATGGCCTGACTGAGGCCATTGCCAGCCATAGGGCCCTGGTGAAGGCCAGGGGCACCAGGCCCTTTGTGATCAGCAGGTCTACCTTTGCTGGCCATGGCAGGTATGCTGGCCACTGGACTGGGGATGTGTGGTCTTCTTGGGAGCAGCTGGCCAGCTCTGTGCCTGAGATCCTGCAGTTCAACCTGCTGGGGGTGCCTCTGGTGGGGGCTGATGTGTGTGGCTTTCTGGGCAACACCTCTGAGGAGCTGTGTGTGAGGTGGACCCAGCTGGGGGCCTTTTACCCCTTCATGAGGAACCACAATAGCCTGCTGAGCCTGCCCCAGGAGCCTTACTCTTTCTCTGAGCCTGCCCAGCAGGCCATGAGGAAGGCCCTGACTCTGAGGTATGCCCTGCTGCCCCATCTGTATACCCTGTTTCACCAGGCCCATGTGGCTGGGGAGACTGTGGCTAGGCCTCTGTTTCTGGAGTTCCCTAAGGACTCTAGCACCTGGACTGTGGACCACCAGCTGCTGTGGGGGGAGGCCCTGCTGATCACCCCTGTGCTGCAGGCTGGCAAGGCTGAGGTGACTGGCTACTTCCCCCTGGGCACCTGGTATGACCTGCAGACTGTGCCTGTGGAGGCCCTGGGGAGCCTGCCTCCCCCCCCTGCTGCCCCCAGGGAGCCTGCCATTCATTCTGAGGGCCAGTGGGTGACCCTGCCTGCCCCTCTGGACACCATCAATGTGCACCTGAGGGCTGGGTACATCATCCCCCTGCAGGGCCCTGGCCTGACCACCACTGAGAGCAGGCAGCAGCCCATGGCCCTGGCTGTGGCTCTGACCAAGGGGGGGGAGGCCAGGGGGGAGCTGTTCTGGGATGATGGGGAGTCTCTGGAGGTGCTGGAGAGGGGGGCCTACACCCAGGTGATCTTTCTGGCCAGGAACAATACTATTGTGAATGAGCTGGTGAGGGTGACCTCTGAGGGGGCTGGCCTGCAGCTGCAGAAGGTGACTGTGCTGGGGGTGGCCACTGCCCCCCAGCAGGTCCTGAGCAATGGGGTGCCTGTGAGCAACTTCACCTACTCTCCTGACACCAAGGTGCTGGACATTTGTGTGTCTCTGCTGATGGGGGAGCAGTTCCTGGTGAGCTGGTGCTGACTCGAGAGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGACTCGAGATCCACTAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO: 25: Amino acid sequence of secreted GAA protein

MAFLWLLSCWALLGTTFGLLVPRELSGSSPVLEETHPAHQQGASRPGPRDAQAHPGRPRAVPTQCDVPPNSRFDCAPDKAITQEQCEARGCCYIPAKQGLQGAQMGQPWCFFPPSYPSYKLENLSSSEMGYTATLTRTTPTFFPKDILTLRLDVMMETENRLHFTIKDPANRRYEVPLETPHVHSRAPSPLYSVEFSEEPFGVIVRRQLDGR VLLNTTVAPLFFADQFLQLSTSLPSQYITGLAEHLSPLMLSTSWTRITLWNRDLAPTPGANLYGSHPFYLALEDGGSAHGVFLLNSNAMDVVLQPSPALSWRSTGGILDVYIFLGPEPKSVVQQYLDVVGYPFMPPYWGLGFHLCRWGYSSTAITRQVVENMTRAHFPLDVQWNDLDYMDSRRDFTFNKDGFRDFPA MVQELHQGGRRYMMIVDPAISSSGPAGSYRPYDEGLLRRGVFITNETGQPLIGKVWPGSTAFPDFTNPTALAWWEDMVAEFHDQVPFDGMWIDMNEPSNFIRGSEDGCPNNELENPPYVPGVVGGTLQAATICASSHQFLSTHYNLHNLYGLTEAIASHRALVKARGTRPFVISRSTFAGHGRYAGHWTGDVWSSWEQLASSVPEILQ FNLLGVPLVGADVCGFLGNTSEELCVRWTQLGAFYPFMRNHNSLLSLPQEPYSFSEPAQQAMRKALTLRYALLPHLYTLFHQAHVAGETVARPLFLEFPKDSSTWTVDHQLLWGEALLITPVLQAGKAEVTGYFPLGTWYDLQTVPVEALGSLPPPPAAPREPAIHSEGQWVTLPAPLDTINVHLRAGYIIPLQGPGLTTTESR QQPMALAVALTKGGEARGELFWDDGESLEVLERGAYTQVIFLARNNNTIVNELVRVTSEGAGLQLQKVTVLGVATAPQQVLSNGVPVSNFTYSPDTKVLDICVSLLMGEQFLVSWC

SEQ ID NO: 26: CpG Reduced Poly-A BGH Nucleic Acid Sequence

AGATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCTCTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAACAATAGCAGGCATG CTGGGGATGCAGTGGGCTCTATGG

SEQ ID NO: 27: Poly-A wtBGH Nucleic Acid Sequence

AGATCTACCGGTGAATTCACCGCGGGTTTAAACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGGCTAGCTCTAGA

SEQ ID NO: 28: ApoE/hAAT sequence flanked by ApaI restriction sites (underlined).

GGGCCC ATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATCTTGCTACCAGTGGAACAGCCACTAAGGATTTCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTC TGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACA GGGCCC

SEQ ID NO: 29: ApoE/hAAT sequence.

ATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGGGATCTTGCTACCAGTGGAACAGCCACTAAGGATCTCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGG TACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACA

AAV vector capsids:

VP1 (SEQ ID NO: 30)

MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDNGRGLVLPGYKYLGPFNGLD

KGEPVNAADAAALEHDKAYDQQLQAGDNPYLRYNHADAEFQERLQEDTSFGGNLGRAVFQ

AKKRVLEPLGLVESPVKTAPGKKRPVEPSPQRSPDSSTGIGKKGQQPAKKRLNFGQTGDS

ESVPDPQPIGEPPAAPSGVGPNTMAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRV

ITTSTRTWALPTYNNHLYKQISNGTSGGSTNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQ

RLINNNWGFRPKRLNFKLFNIQVKEVTQNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSA

HQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFEFSYNFED

VPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQSTGGTAGTQQLLFSQAGPNNMSAQAKNW

LPGPCYRQQRVSTTLSQNNNSNFAWTGATKYHLNGRDSLVNPGVAMATHKDDEERFFPSS

GVLMFGKQGAGKDNVDYSSVMLTSEEEIKTTNPVATEQYGVVADNLQQQNAAPIVGAVNS

QGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADP

PTTFNQAKLASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTE

GTYSEPRPIGTRYLTRNL

VP2 (SEQ ID NO: 31)

TAPGKKRPVEPSPQRSPDSSTGIGKKGQQPAKKRLNFGQTGDSESVPDPQPIGEPPAGPS

GLGSGTMAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHL

YKQISNGTSGGSTNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFK

LFNIQVKEVTQNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMIPQ

YGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFEFSYNFEDVPFHSSYAHSQSLDRLM

NPLIDQYLYYLSRTQSTGGTAGTQQLLFSQAGPNNMSAQAKNWLPGPCYRQQRVSTTLSQ

NNNSNFAWTGATKYHLNGRDSLVNPGVAMATHKDDEERFFPSSGVLMFGKQGAGKDNVDY

SSVMLTSEEEIKTTNPVATEQYGVVADNLQQQNAAPIVGAVNSQGALPGMVWQNRDVYLQ

GPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFNQAKLASFITQYS

TGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTEGTYSEPRPIGTRYLTRN

L

VP3 (SEQ ID NO: 32)

MAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQISN

GTSGGSTNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQV

KEVTQNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTL

NNGSQAVGRSSFYCLEYFPSQMLRTGNNFEFSYNFEDVPFHSSYAHSQSLDRLMNPLIDQ

YLYYLSRTQSTGGTAGTQQLLFSQAGPNNMSAQAKNWLPGPCYRQQRVSTTLSQNNNSNF

AWTGATKYHLNGRDSLVNPGVAMATHKDDEERFFPSSGVLMFGKQGAGKDNVDYSSVMLT

SEEEIKTTNPVATEQYGVVADNLQQQNAAPIVGAVNSQGALPGMVWQNRDVYLQGPIWAK

IPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFNQAKLASFITQYSTGQVSV

EIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTEGTYSEPRPIGTRYLTRNL

LK03-AAV VP1 (SEQ ID NO: 33)

MAADGYLPDWLEDNLSEGIREWWALQPGAPKPKANQQHQDNARGLVLPGYKYLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVDQSPQEPDSSSGVGKSGKQPARKRLNFGQTGDSESVPDPQ PLGEPPAAPTSLGSNTMASGGGAPMADNNEGADGVGNSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISSQSGASNDNHYFGYSTPWGYFDF NRFHCHFSPRDWQRLINNNWGFRPKKLSFKLFNIQVKEVTQNDGTTTIANNLTSTVQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMVPQYGYLTLNN GSQAVGRSSFYCLEYFPSQMLRTGNNFQFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLNRTQGTTSGTTNQSRLLFSQAGPQSMSLQARNWLP GPCYRQQRLSKTANDNNNSNFPWTAASKYHLNGRDSLVNPGPAMASHKDDEEKFFPMHGNLIFGKEGTTASNAELDNVMITDEEEIRTTNPVATEQYG TVANNLQSSNTAPTTRTVNDQGALPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPPPQIMIKNTPVPANPPTTFSPAKFASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYNKSVNVDFTVDTNGVYSEPRPIGTRYLTRPL

EXAMPLE 2

Research design - activity in vitro (Huh7 cells)

[0410] To evaluate the activity of the GAA expression cassettes, 200 ng of plasmid DNA corresponding to each cassette was transfected into a human hepatocellular carcinoma cell line (Huh7) using Lipofectamine® 2000 (ThermoFisher). After 48 hours of incubation, culture medium samples of cells transfected with plasmids carrying each expression cassette were assessed for GAA levels by GAA activity assay and normalized for intracellular protein content.

results

[0411] Figure 10 shows the in vitro evaluation of various GAA expression plasmids in Huh7 cells by GAA activity assay. Results represent the average of three biological parallels n=3. Error bars correspond to standard deviation.

[0412] With the exception of GAA5.wtBGH, all plasmids showed increased expression levels of secreted GAA compared to the control green fluorescent protein (GFP) plasmid, indicating that these plasmids resulted in expression of the GAA protein product (Figure 10). In general, “BGH” plasmids (carrying a CpG-reduced poly-A sequence) were superior to “wtBGH” plasmids (carrying wild-type bGH poly-A). In addition, the codon-optimized sequences of GAA2, GAA7, GAA8, and GAA13 appeared to be relatively similar and tended to be more highly expressed from GAA13 plasmids. An overall decrease in expression levels was observed compared to the parental spΔGAApar plasmid.

EXAMPLE 3

Study Design - Cassette Activity by Hydrodynamic Injection in Mice

[0413] To evaluate the activity of different GAA expression cassettes in mammals, 25 micrograms of plasmid DNA corresponding to each cassette was administered to male C57BL/6 mice (approximately 8 weeks old) intravenously into the lateral tail vein by hydrodynamic injection. Plasma was collected 72 hours after injection and assessed for the presence of circulating GAA enzyme by GAA activity assay and Western blotting.

results

[0414] Figures 11 and 12 show the evaluation of different GAA expression cassettes by hydrodynamic injection into male C57BL/6 mice by GAA activity assay and Western blot, respectively, in plasma. As shown in Figure 11, plasma GAA levels were normalized to the parent construct (spΔGAApar). Results represent the average of n=4-5 biological parallels. Error bars correspond to standard deviation. NS, not significant. * p<0.05, ** p<0.01.

[0415] As shown in Figure 12, in the Western blot analysis of plasma GAA, the bar graph represents the quantitative analysis of the top band normalized to the bottom, non-specific band. Each biological parallel is shown (n=4-5 per group).

[0416] Evaluation of codon-optimized GAA expression cassettes in mice by hydrodynamic injection showed that these cassettes can result in expression and secretion of the GAA transgene product from mouse liver at levels similar to those observed with the parent GAA spΔGAApar construct (Figure 11). As in the in vitro studies on Huh7 cells (Example 2), GAA-expressing cassettes carrying the CpG-reduced poly-A BGH sequence were superior to wild-type poly-A BGH cassettes. Surprisingly, compared to the results in Example 2, some of the codon-optimized GAA constructs performed as well or better than the parent spΔGAApar plasmid.

EXAMPLE 4

Study Design - Vector Activity in Mice

[0417] To evaluate the activity of the codon-optimized GAA cassettes of the invention compared to the parent spΔGAApar cassette, spΔGAApar and 9 codon-optimized cassettes were packaged into an AAV vector containing capsids SEQ ID NO: 30-32 (Table 1). Five male C57BL/6 mice per group were injected intravenously via the tail vein with 1 of 2 doses of vector (5 x 10 ¹¹ VG/kg or 2 x 10 ¹² VG/kg). Circulating GAA activity was measured over time after administration of the AAV vector to mice. Plasma was tested for GAA activity every two weeks. On day 70, 7 of the 10 groups of test animals were sacrificed, and the remaining three groups (SPK-AAV-01, SPK-AAV-02 and parental vector) were sacrificed on day 147.

results

[0418] Figure 13 shows the evaluation of various codon-optimized GAA expression cassettes packaged in an AAV vector (SEQ ID NO: 30-32) in male C57BL/6 mice by plasma GAA activity assay 70 days after vector infusion . Results represent the average of n=4-5 biological parallels. Error bars correspond to standard deviation. NS, not significant. * p<0.05, ** p<0.01.

[0419] Two vectors, SPK-AAV-01 and SPK-AAV-02, consistently demonstrated significantly higher plasma GAA activity (approximately 2-3 times higher at high dose) compared to the parent vector (referred to herein as as SPK-AAV-11).

[0420] Figure 14 shows the evaluation of various codon-optimized GAA expression cassettes packaged in an AAV vector (SEQ ID NO: 30-32) in male C57BL/6 mice by Western blotting of plasma GAA 7 days postinfusion vector. Results represent the average of n=4-5 biological parallels. Error bars correspond to standard deviation. NS, not significant. * p<0.05, ** p<0.01.

[0421] Figure 15 shows GAA activity levels measured in plasma after administration of codon-optimized GAA expression cassettes packaged in an AAV vector (SEQ ID NO: 30-32) to mice. Male C57BL/6 mice were injected with a dose of 5 x 10 ¹¹ (upper panel) or 2 x 10 ¹² VG/kg (lower panel) of either of the two best performing GAA-expressing AAV vectors (SEQ ID NO: 30-32) (n= 5 animals/group). For comparison, GAA activity measured in the plasma of mice injected with the SPK-AAV-11 vector (spΔGAApar parent construct) is shown. From day 42 onwards, plasma GAA activity was significantly higher for the SPK-AAV-01 and SPK-AAV-02 vectors than for the SPK-AAV-11 vector (***p≤0.001,****p≤0. 0001 by Tukey-Kramer multiple comparison test).

EXAMPLE 5

Study Design - Activity in Rats

[0422] The purpose of this study was to evaluate the activity of SPK-AAV-02 in two different strains of rats. Circulating GAA activity was measured in plasma after vector administration to Wistar Hannover rats (n=4) and Sprague-Dawley rats (n=5). Male mice were injected intravenously through the tail vein with 2×10 ¹³ VG/kg vector.

results

[0423] Figure 16 shows the evaluation of SPK-AAV-02 (pAAV-ApoE/hAAT.GAA13.BGH packaged in an AAV vector (SEQ ID NO: 30-32)) in male Wistar Hanover rats and Sprague-Dawley rats compared with C57BL/6 mice using a plasma GAA activity assay. Results represent the average of n=4-5 biological parallels. Error bars correspond to standard deviation.

[0424] GAA activity levels began to plateau on day 28 with measured GAA activity of approximately 15,000 nmol/h/ml. Mice were injected with SPK-AAV-02 at a 10-fold lower dose (2×10 ¹² VG/kg vector) than rats.

EXAMPLE 6

Study Design - Activity in Non-Human Primates (NHP), Part 1

[0425] To evaluate the long-term effects of secreted GAA expression, a single dose of vector SPK-AAV-01, SPK-AAV-02, or SPK-AAV-10 was administered to a species phylogenetically related to humans, rhesus monkeys ( Macaca mulatta ), and the animals were assessed for Plasma GAA activity levels.

[0426] Male rhesus monkeys (n=3/dose cohort) were given a 30-minute IV infusion into the saphenous vein of the leg at a single dose of 6×10 ¹² VG/kg vector SPK-AAV-01, SPK-AAV-02 or SPK- AAV-10. Following administration of the AAV vector, animals were monitored for clinical parameters daily and blood was collected weekly.

results

[0427] Rhesus macaques were injected with a dose of 6×10 ¹² VG/kg GAA-expressing AAV vectors (SEQ ID NO: 30-32) (n=3 animals/group). Figure 17 shows the levels of GAA activity measured in plasma after administration of GAA expression cassettes packaged in an AAV vector (SEQ ID NO: 30-32) to male rhesus monkeys. No significant differences in plasma GAA activity levels were observed in NHPs injected with the SPK-AAV-01, SPK-AAV-02, and SPK-AAV-10 vector.

[0428] Preliminary data from the NHP study confirm the results observed in the mouse studies and suggest that plasma GAA activity levels resulting from administration of SPK-AAV-01 and SPK-AAV-02 are similar (Figure 17) .

EXAMPLE 7

Study Design - Activity in Non-Human Primates, Part 2

[0429] The primary objective of this study was to evaluate the transduction profile of SPK-AAV-01 and SPK-AAV-02 in vervet monkeys (AGM; Chlorocebus sabaeus ). AGM males (n=4/dose cohort) received a 30-minute IV infusion into the saphenous vein of the leg at a single dose of 6x10 ¹² VG vector/kg SPK-AAV-01 or SPK-AAV-02. Following administration of the AAV vector, animals were monitored for clinical parameters daily and blood was collected weekly.

results

[0430] Preliminary data in the vervet monkey study confirm the results observed in mouse studies that plasma GAA activity levels resulting from administration of SPK-AAV-01 and SPK-AAV-02 are similar (Figure 18).

EXAMPLE 8

conclusions

[0431] Codon-optimized GAA cassettes expressing secreted human GAA represent a significant improvement over the parental spΔGAApar construct. In particular, SPK-AAV-01 and SPK-AAV-02 demonstrate exceptional activity and expression of secreted human GAA in cell culture, rodents, and two non-human primate models.

EXAMPLE 9

Study Design - Dose Ranges in NHPs

[0432] To evaluate the dose effect in the case of expression of secreted GAA, three doses of the SPK-AAV-02 vector were administered to a primate species phylogenetically related to humans, rhesus monkeys ( Macaca mulatta ), and the animals were assessed for levels of GAA activity over one month.

[0433] Male rhesus monkeys (n=4/dose cohort) were given a 30-minute intravenous (IV) infusion into the saphenous vein of the leg at a dose of 2 x 10 ¹² , 6 x 10 ¹² , 2 x 10 ¹³ VG/kg SPK- vector AAV-02. Female rhesus monkeys (n=4/dose cohort) received a 30-minute IV infusion into the saphenous vein of the leg at a dose of 2x10 ¹³ VG/kg SPK-AAV-02 vector. Following administration of the AAV vector, animals were monitored for clinical signs daily. Blood was collected weekly for 4 weeks to measure GAA activity, GAA antigen levels, anti-GAA IgG production, complete blood count and biochemistry, including glucose levels, to monitor potential changes in glycemia.

results

[0434] Figure 21 shows GAA activity levels measured in plasma following administration of the SPK-AAV-02 vector to male and female rhesus monkeys. Results for GAA activity measured in plasma 7, 14, 21 and 28 days after vector injection indicate a correlation between vector dose and plasma GAA activity level. Levels of GAA activity in the plasma of female NHPs injected with a dose of 2x10 ¹³ VG/kg were approximately 0.5 times lower (49.6%) than those measured in male NHPs injected with an equal dose.

[0435] The data in this NHP study confirm the results observed in mouse studies, namely that plasma GAA activity levels achieved using AAV vectors to express secreted GAA result in a dose-dependent plasma GAA activity following vector administration (Figure 21). Following a single infusion of SPK-AAV-02 NHP at three escalating doses, dose-dependent expression of GAA in NHP plasma was observed.

EXAMPLE 10

NHP study for nine months

[0436] Each of SPK-AAV-01, SPK-AAV-02 and SPK-AAV-10 was administered in a single dose (6×10 ¹² VG/kg) via intravenous infusion to male rhesus monkeys ( Macaca mulatta ). During the study, a subset of animals injected with vectors expressing secreted GAA exhibited a decrease in active plasma GAA protein and a concomitant increase in levels of anti-GAA IgG antibodies. The loss of GAA activity and plasma antigen levels likely resulted from the development of IgG-mediated humoral immune clearance of the protein product of the human transgene. To reduce the humoral immune response and possibly restore detectable levels of circulating GAA enzyme, starting at day 183, an immunosuppressive treatment regimen of monthly rituximab in combination with daily cyclosporine A was used in some animals.

results

[0437] Plasma levels of secreted and active GAA were determined in all animals, observing peak mean activity levels per day of 15,482.07 ± 47.90, 664.43 ± 417.55, and 427.833 ± 307.94 nmol/ml/h in case SPK-AAV-01, SPK-AAV-02 and SPK-AAV-10, respectively. Detectable levels of circulating GAA antigen were observed with mean peak levels of 14.84±2.21, 19.11±23.23, 14.64±16.96 ng/ml in animals administered SPK-AAV-01, SPK-AAV -02 and SPK-AAV-10, respectively. Concomitant with the observed decrease in circulating GAA, an increase in circulating anti-GAA IgG levels was observed from day 15 onwards. Using qPCR analysis of vector genomes in liver tissue, the presence of a vector in the liver was confirmed, corresponding to an average number of vector genome copies of 6.39 ± 2.72, 11.43 ± 4.23 and 19.01 ± 13.93 per haploid genome, in case of animals administered SPK-AAV-01, SPK-AAV-02 and SPK-AAV-10, respectively.

[0438] Liver samples were analyzed for levels of full-length GAA and cleaved GAA (lysosomal form). The test samples were compared to liver samples from vector-naïve rhesus macaques serving as negative controls that did not exhibit detectable levels of cleaved or intact GAA at protein loading levels (n=3). The amount of full-length protein in the liver of the studied animals was 4.0±7.0, 5.4±9.1, 11.0±11.3 ng GAA/mg total protein in animals that were administered SPK-AAV-01, SPK- AAV-02 and SPK-AAV-10, respectively. Each cohort contained higher levels of cleaved GAA than full-length GAA, with levels of 41.6 ± 57.6, 44.4 ± 74.5, 49.0 ± 40.3 ng GAA/mg total protein observed in animals fed SPK-AAV-01, SPK-AAV-02 and SPK-AAV-10 were administered, respectively. Regardless of GAA processing, differences between groups were not significant (P=0.72 for full-length (intact) form; P=0.83 for lysosomal (cleaved) form by Kruskal-Wallis test). Finally, all groups administered the AAV vectors of the invention exhibited mean detectable liver GAA levels above baseline levels.

conclusions

[0439] Administration of each of SPK-AAV-01, SPK-AAV-02 and SPK-AAV-10 resulted in detectable and elevated levels of GAA antigen in plasma, peaking at weeks 2-3 and linearly correlating with GAA activity, wherein similar kinetics are observed regardless of the vector genome. GAA activity and plasma GAA antigen levels decreased consistently at subsequent time points in all but one animal. As previously observed in NHPs injected with AAV vectors, a humoral response to the human transgene product was observed beginning at day 43 that was inversely correlated with plasma GAA antigen levels in all animals that developed a GAA-specific IgG response. Suppression of anti-GAA IgG levels by administration of an immunosuppressive agent targeting the B-cell mediated IgG response (rituximab) resulted in the restoration of detectable plasma GAA levels in a subset of animals. In all groups, copies of the vector genome were detectable in the liver at the end of the study, even in the absence of measurable levels of circulating GAA in plasma. Accordingly, GAA protein was detectable in all liver tissues by the end of the study in the case of vector administration, while GAA antigen was below the detection limit in unexposed liver tissue. Taken together, these results indicate that the AAV vectors of the invention can mediate long-lasting expression of secreted human GAA in the liver for more than 9 months, independent of the development of a humoral response to the human transgene product.

EXAMPLE 11

Nine-week immunosuppression study in rodents

[0440] Expression levels of the GAA transgene product were measured for nine weeks after intravenous administration of a single dose of 2×10 ¹² VG/kg SPK-AAV-02 particles in the presence or absence of rapamycin and/or prednisolone in male and female C57BL/6 mice. Mice were tested in 5 groups as shown in Table 2.

Table 2 - Definition of groups and dose levels

Group Number of animals SPK-AAV-02 (VG/kg) Rapamycin
(2-3 mg/kg)* Prednisolone
(1-0.25 mg/kg)** Males Females 1 5 5 Carrier 2 10 10 2×10 ¹² 3 10 10 X 4 10 10 X 5 10 10 X X

VG=vector genomes, X=introduction;

* Rapamycin was administered daily at 2 mg/kg from day (-)7 to day 5. Beginning on day 6, administration was changed to 3 mg/kg every other day;

** Prednisolone was administered at a concentration of 1 mg/kg from day (-)1 to day 15, 0.5 mg/kg from day 16 to day 22, and 0.25 mg/kg from day 23 to day 28.

[0441] In the groups that received immunosuppressive agents, the agents were administered before administration of AAV particles and during the first 5 weeks of the study. After completion of AAV administration, animals were monitored for clinical parameters and body weight for 9 weeks. At the end of the study, clinical and biochemical blood tests and histopathological analysis of selected tissues were performed. IgG antibodies against the rAAV capsid and plasma GAA activity were assessed weekly throughout the study.

results

[0442] Plasma GAA activity levels above background levels were determined in all vector-treated animals. In vehicle-treated males in group 1, background levels of approximately 300-400 nmol/ml/h were observed. Circulating GAA activity levels increased following vector administration with slightly different kinetics depending on the immunosuppressive treatment group, but showed equivalent peak mean GAA activity levels. Specifically, the peak mean GAA activity levels were 16114±5411, 14875±6882, 14890±6882 and 21480±6340 nmol/ml/h for males in group 2 (SPK-AAV-02 alone), group 3 (SPK- AAV-02, 2-3 mg/kg rapamycin), group 4 (SPK-AAV-02, 1-0.25 mg/kg prednisolone) and group 5 (SPK-AAV-02, 2-3 mg/kg rapamycin, 1-0.25 mg/kg prednisolone), respectively. No statistically significant differences in peak GAA activity levels were observed between any of the AAV administration groups.

[0443] In female mice, peak mean GAA activity levels were 7380±4034, 5912±3259, 6096±3249 and 9955±3104 nmol/ml/h in group 2, group 3, group 4 and group 5, respectively. These levels of GAA activity were significantly higher than background levels detected in vehicle-treated female animals. As with male mice, peak GAA activity levels in female mice were not statistically significantly different between any of the AAV treatment groups. The observed differences in plasma GAA activity levels between males and females are consistent with the well-described phenomenon of reduced AAV transduction of hepatocytes in female mice (Davidoff et al ., 2003, Blood, 102:480-488; DOI: 10.1182/blood-2002-09-2889) .

[0444] Plasma anti-rAAV capsid IgG was determined in all SPK-AAV-02-treated animals, observing peak mean levels of 86527±92140, 6695±3555, 64368±29635 and 11374±6053 ng/ml in males in group 2. group 3, group 4 and group 5, respectively, and peak mean levels of 182009±148148, 66141±77925, 182654±90161 and 57752±59192 ng/ml in females in groups 2, 3, 4 and 5, respectively. In group 1 (vehicle), males and females had anti-rAAV capsid IgG levels below the quantitative limit of the assay at all time points.

Conclusion

[0445] Administration of SPK-AAV-02 resulted in detectable levels of GAA activity in all animals. Combination treatment with rapamycin and prednisone resulted in GAA activity levels that were statistically elevated (compared to rAAV vector alone) on days 22, 36, and 42 in male mice and on day 43 in female animals. It should be noted that differences in peak levels of GAA activity, regardless of when maximum expression was achieved, were not statistically significant between any of the AAV administration groups among the sexes.

[0446] A humoral IgG response induced against the rAAV capsid was observed in all groups administered SPK-AAV-02. In both sexes, the presence of rapamycin (groups 3 and 5) significantly reduced the production of IgG against the capsid compared with the vector alone. Prednisolone treatment had a more modest effect on anti-capsid IgG production, with a significant decrease at days 15, 43 and 60 in males and days 43 and 60 in females in group 4.

[0447] Immunosuppression with rapamycin and/or prednisolone in mice resulted in reduced humoral responses to the rAAV capsid and did not significantly affect peak plasma GAA activity levels.

EXAMPLE 12

4-Week Single-Dose Study of NHP Coadministered with Rapamycin

[0448] This study assessed circulating GAA activity (plasma GAA activity levels) over a period of four weeks following administration of a single dose of SPK-AAV-02 5.5 x 10 ¹³ VG/kg in the presence of rapamycin to three male vervet monkeys ( Chlorocebus sabaeus ). .

[0449] Animals were administered rapamycin (2 mg/kg daily) starting 5 days before AAV administration and continuing throughout the four-week study. A single dose of 5.5×10 ¹³ VG/kg SPK-AAV-02 was administered intravenously through the saphenous vein of the leg. After completion of AAV administration, animals were monitored for clinical parameters and body weight for 4 weeks. At the end of the study, clinical and biochemical blood tests and histopathological analysis of selected tissues were performed. Plasma GAA activity was measured weekly throughout the study.

results

[0450] Plasma GAA activity levels exceeded background levels in all animals following administration of SPK-AAV-02, with peak activity levels in each of the three animals reaching 1158.1, 711.9, and 623.8 nmol/ml/h at terminal measurement point (Table 3).

Table 3 - Plasma GAA activity in individual NHPs treated with rapamycin.

Plasma GAA activity (nmol/ml/h) Individual NHP numbers 1 2 3 Mean ± SD Baseline 2.1 4.9 3.7 3.6±1.4 Day 7 777.2 331.3 187.4 432.0±307.5 Day 14 924.0 349.9 368.0 547.3±326.4 Day 21 973.2 289.6 490.0 584.3±351.4 Day 28 1158.1 711.9 623.8 831.3±286.5

conclusions

[0451] Administration of SPK-AAV-02 resulted in detectable GAA activity in the plasma of all three NHPs. The combination of SPK-AAV-02 and rapamycin was well tolerated and had a favorable safety profile in all tests performed.

EXAMPLE 13

Therapeutic efficacy in a mouse model of Pompe disease Gaa ^-/-

[0452] A 10-month follow-up study was conducted in 4-month-old Gaa ^-/- mice (a common mouse model of Pompe disease) that were administered SPK-AAV-02 at three doses (1.25 x 10 ¹¹ , 5 x 10 ¹¹ and 2×10 ¹² VG/kg). The study showed: (1) a dose-dependent increase in circulating GAA enzyme activity, (2) a significant improvement in muscle strength, and (3) the absence of significant humoral immune responses against GAA.

EXAMPLE 14

Methods

Cell-based assay to measure protein expression efficiency from plasmids encoding human secreted GAA transgenes

[0453] Huh7 cells were seeded in 48-well plates at 5 x 10 ⁴ cells per well overnight in DMEM+10% FBS+penicillin/streptomycin/L-glutamine. Using plasmids prepared using the Plasmid Giga Kit (Qiagen), cells were transfected at 250 ng per well using Lipofectamine ^® 2000. Cells were maintained at 37°C and 5% CO ₂ for 48-72 hours and supernatants were collected and were stored in low-retention microtiter plates at −80°C until assayed for GAA activity. Supernatants were diluted 1:10 and incubated with 3 mM fluorescent substrate 4-methylumbelliferyl-α-D-glucopyranoside for 1 hour at 37°C. The reaction was stopped after 1 hour with carbonate buffer (pH 10.5) and compared with a standard curve obtained by diluting 4-methylumbelliferone (4-MU) in stop buffer and generating a 12-point standard curve starting at 250 pmol/μl and ending at 0 pmol/µl. The plate was read at λex 360 nm; λem 449 nm.

Hydrodynamic injection of plasmid DNA to assess the activity of plasmids encoding human secreted GAA transgenes

[0454] Plasmids prepared using the Plasmid Giga Kit (Qiagen) were diluted in PBS180+0.001% Pluronic. 25 μg of plasmid DNA corresponding to each cassette to be tested was administered to male C57BL/6 mice approximately 8 weeks old intravenously into the lateral tail vein by hydrodynamic injection. 72 hours after injection, whole blood was collected into sodium citrate tubes by venesection of the submandibular vein. The first drop of blood was discarded, 200 μl of blood was collected in sodium citrate and the tube was inverted to avoid hemolysis and clotting. The sample was centrifuged at 10,000 rpm for 10 minutes at 4°C. Plasma was aliquoted into tubes and stored in a freezer at -80°C until assayed for GAA activity. Plasma samples were diluted to varying degrees and incubated with 3 mM fluorescent substrate 4-methylumbelliferyl-α-D-glucopyranoside for 1 hour at 37°C. The reaction was stopped after 1 hour with carbonate buffer (pH 10.5) and compared to a standard curve obtained by diluting 4-MU in stop buffer and generating a 12-point standard curve starting at 250 pmol/μl and ending at 0 pmol/ ml. The plate was read at λex 360 nm; λem 449 nm.

Cell-based assay to measure the activity of AAV vectors encoding human secreted GAA transgenes

[0455] Huh7 cells were seeded in 48-well plates at 5 x 10 ⁴ cells per well overnight. SPK-AAV-01 and SPK-AAV-02 from the stock vector were prepared according to the dose curve (multiplicity of infection, MOI, range 2×10 ⁶ -1×10 ⁴ ) in Opti-MEM ^TM or DMEM+10% FBS+penicillin /streptomycin/L-glutamine. Existing media was removed from Huh7 cells and replaced with media containing viral particles. Cells were maintained at 37°C and 5% CO ₂ for 72 hours, and supernatants were collected and stored in low-retention microtiter plates at -80°C until assayed for GAA activity. Supernatants were diluted 1:10 and incubated with 3 mM fluorescent substrate 4-methylumbelliferyl-α-D-glucopyranoside for 1 hour at 37°C. The reaction was stopped after 1 hour with carbonate buffer (pH 10.5) and compared to a standard curve obtained by diluting 4-MU in stop buffer and generating a 12-point standard curve starting at 250 pmol/μl and ending at 0 pmol/ µl The plate was read at λex 360 nm; λem 449 nm.

Analysis of GAA activity in mouse/rat plasma

[0456] Whole blood was collected into sodium citrate tubes by venesection of the submandibular vein. The first drop of blood was discarded and 200 μl of blood was collected in sodium citrate; the tube was inverted to avoid hemolysis and clotting. The sample was centrifuged at 10,000 rpm for 10 minutes at 4°C. Plasma was aliquoted into tubes and stored in a -80°C freezer until analyzed for GAA activity. Plasma samples were diluted to varying degrees and incubated with 3 mM fluorescent substrate 4-methylumbelliferyl-α-D-glucopyranoside for 1 hour at 37°C. The reaction was stopped after 1 hour with carbonate buffer (pH 10.5) and compared to a standard curve obtained by diluting 4-MU in stop buffer and generating a 12-point standard curve starting at 250 pmol/μl and ending at 0 pmol /µl The plate was read at λex 360 nm; λem 449 nm.

GAA Activity Assay in NHP (Rhesus/AGM) Plasma

[0457] Male rhesus/green monkeys were given an intravenous injection through the saphenous vein of the leg and whole blood was collected into sodium citrate tubes, which were inverted to avoid hemolysis and clotting. The sample was centrifuged at 10,000 rpm for 10 minutes at 4°C. Plasma was aliquoted into tubes and stored in a freezer at -80°C until assayed for GAA activity. Plasma samples were diluted to varying degrees and incubated with 3 mM fluorescent substrate 4-methylumbelliferyl-α-D-glucopyranoside for 1 hour at 37°C. The reaction was stopped after 1 hour with carbonate buffer (pH 10.5) and compared to a standard curve obtained by diluting 4-MU in stop buffer and generating a 12-point standard curve starting at 250 pmol/μl and ending at 0 pmol/ µl, or 11-point standard curve (5 µM/ml - 0.49 nM/ml). The plate was read at λex 360 nm; λem 449 nm. GAA activity for test samples was interpolated from the 4-MU standard curve and expressed as rate (nanomole/milliliter/hour (nmol/ml/h)).

Assessment of GAA protein levels in Huh7 supernatants by Western blotting

[0458] Huh7 cells were seeded in 48-well plates at 5 x 10 ⁴ cells per well overnight. SPK-AAV-01 and SPK-AAV-02 from stock vector were prepared according to the dose curve (multiplicity of infection, MOI, range 2×10 ⁶ -1×10 ⁴ ) in Opti-MEM ^TM or DMEM+10% FBS+penicillin /streptomycin/L-glutamine. Existing media was removed from Huh7 cells and replaced with media containing viral particles. Cells were maintained at 37°C and 5% CO ₂ for 72 hours, and supernatants were collected and stored in low-retention microtiter plates at -80°C until analyzed for GAA protein levels. Supernatants were diluted 1:20 with RIPA buffer, incubated at 95°C for 5 minutes, and analyzed on a NuPage 4–12% bis-Tris gel with MOPS running buffer. Protein was transferred to a polyvinylidene difluoride (pvdf) membrane using the iBlot ^® system (ThermoFisher) and blocked for 1 hour at room temperature in Odyssey ^® TBS buffer. The membrane was incubated overnight at 4°C with the primary antibody, rabbit anti-human GAA antibody (Abcam), diluted 1:1000. The membrane was washed and incubated at room temperature for 2 hours with fluorescently labeled anti-rabbit secondary antibody diluted 1:10,000. The membrane was washed and imaged using the Li-Core ^® imaging system. Bands were compared with 10 ng Myozyme ^® and marker. Densitometric analysis was performed using Li-Core ^® software and bands were normalized to the Myozyme ^® control.

Assessment of GAA protein levels in mouse plasma by Western blotting

[0459] Male and female C57BL/6 mice were given an intravenous injection via the tail vein and whole blood was collected into sodium citrate tubes via submandibular vein venesection. The first drop of blood was discarded and 200 μl of blood was collected in sodium citrate; the tube was inverted to avoid hemolysis and clotting. The sample was centrifuged at 10,000 rpm for 10 minutes at 4°C. Plasma was aliquoted into tubes and stored in a freezer at -80°C until GAA protein levels were analyzed. Plasma was diluted 1:20 with RIPA buffer, incubated at 95°C for 5 minutes, and analyzed on a NuPage 4–12% bis-Tris gel with MOPS running buffer. Protein was transferred to pvdf membrane using the iBlot ^® system (ThermoFisher) and blocked for 1 hour at room temperature in Odyssey ^® TBS buffer. The membrane was incubated overnight at 4°C with the primary antibody, rabbit anti-human GAA antibody (Abcam), diluted 1:1000. The membrane was washed and incubated at room temperature for 2 hours with fluorescently labeled anti-rabbit secondary antibody diluted 1:10,000. The membrane was washed and imaged using the Li-Core ^® imaging system. Bands were compared with 10 ng Myozyme ^® and marker. Densitometric analysis was performed using Li- ^Core® software and bands were normalized to the ^Myozyme® control.

Assessment of GAA protein levels in NHP plasma by WES

[0460] Male rhesus/green monkeys were given an intravenous injection through the saphenous vein of the leg and whole blood was collected into sodium citrate tubes, which were inverted to avoid hemolysis and clotting. Samples were centrifuged at 10,000 rpm for 10 minutes at 4°C. Plasma was aliquoted into tubes and stored in a −80°C freezer until analyzed for GAA protein levels. Plasma samples were diluted 1:6000 in sample diluent. An 8-point 2-fold standard curve was added to each ^Myozyme® chip starting at 50 ng/mL. Samples were diluted in buffer, denatured at 95°C for 5 minutes and loaded onto a WES™ chip (Protein Simple). The primary antibody was added at a 1:1000 dilution, and the secondary antibody was added at a working dilution. The chip was analyzed using WES™ Protein Simple software and the resulting bands were analyzed and compared to the Myozyme ^® standard curve and normalized to the non-specific second band.

IgG antibodies against AAV capsid

[0461] Total IgG production against the AAV capsid was measured using a capture ELISA assay. The wells of the ELISA plate were coated with 50 μl of a solution containing 1 μg/ml rAAV particles. Total human IgG (Southern Biotech, 0150-01) was diluted to produce a 10-point standard curve ranging from 10,000 ng/ml to 0.5 ng/ml and added to the plate. The limit of quantitation of the assay was 460 ng/mL after backcalculation. Three levels of quality control samples were prepared and included in each plate to evaluate assay performance. Capsid particles, standards, and quality control samples were incubated overnight at 4°C. After washing, wells were blocked with 2% BSA, 0.05% Tween-20 in PBS for 2 hours at room temperature. Serial dilutions of samples in blocking buffer were applied to the plate and incubated at room temperature for 2 hours. The detector antibody was HRP-conjugated sheep anti-human IgG (GE Healthcare, NA933V) diluted 1:5000 in blocking buffer and incubated in the plate for 1 hour at room temperature. After washing, peroxidase activity was analyzed by incubating it for 10 minutes at room temperature with the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The reaction was stopped with 1 M sulfuric acid and the plate was read using an absorbance spectrophotometer to read optical density (OD) at 450 nm. The IgG concentration was determined relative to a standard curve obtained using serial dilutions of purified total human IgG.

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LIST OF SEQUENCES

<110> SPARK THERAPEUTICS, INC.

ANGUELA, Xavier

ARMOR, Sean

NORDIN, Jayme

<120> CODON-OPTIMIZED ACIDIC EXPRESSION CASSETTES

α-GLUCOSIDASES AND METHODS OF THEIR APPLICATION

<130> 023637-0504752

<140> PCT/US2019/032502

<141> 2019-05-15

<150> 62/672,419

<151> 2018-05-16

<150> 62/734,454

<151> 2018-09-21

<160> 33

<170> Patent In version 3.5

<210> 1

<211> 2808

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 1

atggctttcc tgtggctgct gagctgctgg gctctgctgg gcaccacctt tgggctgctg 60

gtgcctaggg agctgtctgg gtctagccct gtgctggagg agactcaccc tgcccatcag 120

caggggggcta gcaggcctgg ccccagggat gctcaggccc accctggcag gcccaggggct 180

gtgcccaccc agtgtgatgt gccccccaac agcaggtttg actgtgcccc tgacaaggcc 240

attacccagg agcagtgtga ggccaggggc tgctgctaca ttccagctaa gcagggcctg 300

caggggggccc agatgggcca gccctggtgc ttcttccccc ccagctatcc tagctataaa 360

ctggagaacc tgagcagctc tgagatgggc tatactgcca ccctgactag gactactccc 420

accttttttc ctaaggatat cctgaccctg aggctggatg tgatgatgga gactgagaac 480

aggctgcact tcactattaa ggaccctgcc aataggaggt atgaagtgcc tctggagact 540

cctcatgtgc actctagggc ccccagcccc ctgtattctg tggagttctc tgaggagccc 600

tttggggtga ttgtgaggag gcagctggat ggcagggtgc tgctgaacac cactgtggcc 660

cccctgttct ttgctgacca gttcctgcag ctgagcacca gcctgcccag ccagtacatc 720

actgggctgg ctgagcatct gagccctctg atgctgagca cctcttggac caggatcacc 780

ctgtggaata gggatctggc ccccacccct ggggctaatc tgtatggctc tcatcccttt 840

tacctggccc tggaggatgg gggctctgcc catggggtgt ttctgctgaa cagcaatgcc 900

atggatgtgg tgctgcagcc ctctcctgcc ctgagctgga ggagcactgg gggcatcctg 960

gatgtgtaca tcttcctggg ccctgagccc aagtctgtgg tccagcagta tctggatgtg 1020

gtgggctacc cctttatgcc cccctattgg ggcctgggct tccacctgtg caggtggggg 1080

tattcttcta ctgctatcac caggcaggtg gtggagaaca tgaccagggc tcacttcccc 1140

ctggatgtgc agtggaatga cctggactat atggactcta ggagggattt caccttcaac 1200

aaggatggct tcagggactt ccctgctatg gtccaggagc tgcatcaggg gggcaggagg 1260

tacatgatga ttgtggaccc tgccatcagc agctctggcc ctgctggcag ctataggccc 1320

tatgatgagg gcctgaggag gggggtgttt atcactaatg aaactgggca gcccctgatt 1380

ggcaaggtgt ggcctggctc tactgccttc cctgacttca ccaaccccac tgctctggcc 1440

tggtgggagg acatggtggc tgagttccat gaccaggtgc cttttgatgg catgtggatt 1500

gacatgaatg agcccagcaa cttcatcagg ggctctgagg atgggtgccc caataatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ccctgcaggc tgccactatt 1620

tgtgccagct ctcaccagtt cctgagcacc cactacaacc tgcacaatct gtatggcctg 1680

actgaggcca ttgccagcca cagggccctg gtgaaggcca ggggcactag gccctttgtg 1740

atctctagaa gcacctttgc tggccatggg aggtatgctg gccactggac tggggatgtg 1800

tggagctctt gggagcagct ggccagctct gtgcctgaga tcctgcagtt caacctgctg 1860

ggggtgcccc tggtgggggc tgatgtgtgt ggcttcctgg gcaacacctc tgaagagctg 1920

tgtgtgaggt ggacccagct gggggccttc taccctttca tgaggaacca caacagcctg 1980

ctgagcctgc ctcaggagcc ttactctttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgaccc tgaggtatgc tctgctgccc cacctgtaca ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc caggcccctg ttcctggagt ttcctaagga tagcagcacc 2160

tggactgtgg accaccagct gctgtggggg gaggccctgc tgattacccc tgtgctgcag 2220

gctggcaagg ctgaggtgac tggctacttc cccctgggca cttggtatga cctgcagact 2280

gtgcctgtgg aagccctggg cagcctgcct cccccccctg ctgcccccag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgcccccc tggacaccat taatgtgcat 2400

ctgaggctg ggtatattat ccccctgcag gggcctgggc tgactaccac tgagagcagg 2460

cagcagccta tggccctggc tgtggctctg actaaggggg gggaggccag gggggagctg 2520

ttctgggatg atggggagag cctggaggtg ctggagaggg gggcctacac ccaggtgatt 2580

ttcctggcca ggaacaacac cattgtgaat gagctggtga gggtgacctc tgagggggct 2640

ggcctgcagc tgcagaaagt gactgtgctg ggggtggcca ctgcccccca gcaggtgctg 2700

agcaatgggg tgcctgtgag caacttcacc tacagccctg acaccaaggt gctggatatt 2760

tgtgtgagcc tgctgatggg ggagcagttc ctggtgagct ggtgctga 2808

<210> 2

<211> 2808

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 2

atggctttcc tgtggctgct gtcttgctgg gccctgctgg ggactacctt tggcctgctg 60

gtgcccagg aactgtctgg ctctagccca gtgctggagg agacccaccc tgcccaccag 120

caggggggctt ctaggcctgg ccccagggat gcccaggccc accctggcag gccaagggct 180

gtgcccaccc agtgtgatgt gccccccaac tctagatttg attgtgcccc tgataaggcc 240

atcacccagg agcagtgtga ggctaggggc tgctgctaca tccctgctaa gcagggcctg 300

caggggggctc agatgggcca gccctggtgc ttcttccccc ccagctatcc ctcttacaag 360

ctggagaatc tgagcagctc tgagatgggc tacactgcca ccctgaccag gactactccc 420

accttcttcc ccaaggacat cctgaccctg aggctggatg tgatgatgga gactgagaac 480

aggctgcatt tcaccatcaa ggatcctgcc aacaggaggt atgaggtgcc tctggagacc 540

ccccatgtgc acagcagggc tccttctccc ctgtactctg tggagttctc tgaggaaccc 600

tttggggtga ttgtgaggag gcagctggat ggcagggtcc tgctgaacac cactgtggcc 660

cccctgttct ttgctgatca gttcctgcag ctgtccactt ctctgcctag ccagtacatc 720

actgggctgg ctgagcacct gagccctctg atgctgagca cctcttggac taggatcacc 780

ctgtggaaca gggacctggc ccccacccct ggggccaacc tgtatggcag ccaccccttc 840

tatctggccc tggaggatgg gggctctgcc catggggtgt tcctgctgaa tagcaatgct 900

atggatgtgg tgctgcagcc cagccctgcc ctgtcttgga ggagcactgg gggcatcctg 960

gatgtgtaca ttttcctggg gcctgagccc aagtctgtgg tgcagcagta cctggatgtg 1020

gtgggctacc ccttcatgcc tccctactgg ggcctgggct tccacctgtg caggtggggc 1080

tacagctcta ctgccatcac caggcaggtg gtggagaata tgaccagggc ccacttcccc 1140

ctggatgtgc agtggaatga cctggactac atggactcta ggagggactt caccttcaat 1200

aaggatggct tcagagactt ccctgccatg gtgcaggagc tgcatcaggg gggcaggagg 1260

tacatgatga ttgtggaccc tgccatcagc tcttctggcc ctgctggctc ttacaggccc 1320

tatgatgagg gcctgaggag gggggtgttc atcaccaatg agactgggca gcccctgatt 1380

gggaaggtgt ggcctggctc tactgccttc cctgacttca ccaatcctac tgccctggcc 1440

tggtgggagg acatggtggc tgagttccat gaccaggtgc cctttgatgg catgtggatt 1500

gacatgaatg agccctctaa tttcatcagg ggctctgagg atggctgccc caacaatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ccctgcaggc tgccaccatc 1620

tgtgctagct ctcaccagtt cctgagcacc cactacaatc tgcataacct gtatggcctg 1680

actgaggcca ttgccagcca cagggccctg gtgaaggcta ggggcaccag gccctttgtg 1740

atttctagga gcacttttgc tggccatggc aggtatgctg ggcactggac tggggatgtg 1800

tggtctagct gggagcagct ggcttcttct gtgcctgaga tcctgcagtt caacctgctg 1860

ggggtgcctc tggtgggggc tgatgtgtgt gggttcctgg gcaacacttc tgaggagctg 1920

tgtgtgaggt ggacccagct gggggccttc taccctttca tgaggaacca caacagcctg 1980

ctgagcctgc cccaggagcc ctacagcttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgaccc tgaggtatgc cctgctgccc cacctgtaca ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc taggcctctg ttcctggagt tccccaagga ctctagcacc 2160

tggactgtgg accaccagct gctgtggggg gaggccctgc tgatcactcc tgtgctgcag 2220

gctgggaagg ctgaggtgac tggctatttc cccctgggca cctggtatga cctgcagact 2280

gtgcctgtgg aggccctggg gagcctgccc cccccccctg ctgcccccag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgcccctc tggataccat caatgtgcac 2400

ctgaggctg gctacatcat tcccctgcag ggccctggcc tgaccactac tgagtctagg 2460

cagcagccca tggccctggc tgtggccctg accaaggggg gggaggctag gggggagctg 2520

ttttgggatg atggggagag cctggaggtg ctggagaggg gggcctacac tcaggtgatc 2580

ttcctggcca ggaacaatac cattgtgaat gagctggtga gggtgacctc tgagggggct 2640

ggcctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgcccccca gcaggtgctg 2700

agcaatgggg tgcctgtgag caacttcacc tatagccctg ataccaaggt gctggatatt 2760

tgtgtgagcc tgctgatggg ggagcagttc ctggtgagct ggtgctga 2808

<210> 3

<211> 2808

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 3

atggctttcc tgtggctgct gtcttgttgg gctctgctgg gcaccacctt tggcctgctg 60

gtgcccaggg agctgtctgg cagcagccct gtgctggagg agacccaccc tgctcatcag 120

caggggggcta gcaggcctgg ccccagggat gcccaggctc accctgggag acccagggct 180

gtgcccactc agtgtgatgt gccccccaac agcaggtttg actgtgctcc tgacaaggct 240

atcacccagg agcagtgtga ggccagggg tgctgctaca ttcctgctaa gcagggcctg 300

caggggggccc agatgggcca gccctggtgc ttcttccccc cctcttatcc cagctataag 360

ctggagaacc tgagcagctc tgagatgggc tacactgcca ccctgaccag gaccactccc 420

accttctttc ccaaggatat tctgactctg aggctggatg tgatgatgga gactgagaac 480

aggctgcact tcactatcaa ggaccctgcc aataggaggt atgaggtgcc cctggagact 540

cctcatgtgc atagcagggc cccttctcct ctgtattctg tggagttctc tgaggagccc 600

tttggggtga ttgtgaggag gcagctggat ggcagggtgc tgctgaacac cactgtggcc 660

cccctgttct ttgctgacca gttcctgcag ctgagcactt ctctgcccag ccagtacatt 720

actgggctgg ctgagcatct gagccccctg atgctgagca cctcttggac caggatcacc 780

ctgtggaaca gggacctggc ccccactcct ggggctaacc tgtatggctc tcaccccttt 840

tacctggccc tggaggatgg gggctctgcc catggggtgt ttctgctgaa cagcaatgct 900

atggatgtgg tgctgcagcc ctctccagcc ctgtcttgga ggagcactgg gggcattctg 960

gatgtgtaca ttttcctggg gcctgaaccc aagtctgtgg tgcagcagta cctggatgtg 1020

gtgggctacc ccttcatgcc cccctattgg gggctggggt ttcacctgtg caggtggggc 1080

tacagcagca ctgccatcac caggcaggtg gtggagaaca tgaccagggc ccatttcccc 1140

ctggatgtgc agtggaatga cctggactac atggatagca ggagggattt caccttcaac 1200

aaggatggct tcagggactt tcctgccatg gtgcaggagc tgcaccaggg gggcaggagg 1260

tatatgatga ttgtggaccc tgctatcagc agctctggcc ctgctggctc ttacaggccc 1320

tatgatgagg gcctgaggag gggggtgttt atcactaatg aaactggcca gcctctgatt 1380

ggcaaggtct ggcctggctc tactgccttc cctgatttta ctaaccccac tgccctggcc 1440

tggtgggagg acatggtggc tgagttccat gatcaggtgc cttttgatgg catgtggatt 1500

gatatgaatg aaccaagcaa cttcatcaga ggctctgagg atggctgccc caacaatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ctctgcaggc tgccaccatt 1620

tgtgctagca gccaccagtt cctgagcacc cactacaatc tgcacaacct gtatggcctg 1680

actgaagcca ttgccagcca tagggccctg gtgaaggcca ggggcactag gccttttgtg 1740

atcagcagga gcacttttgc tggccatggc aggtatgctg gccactggac tggggatgtg 1800

tggagcagct gggagcagct ggccagctct gtgcctgaga ttctgcagtt taacctgctg 1860

ggggtgcccc tggtgggggc tgatgtgtgt ggcttcctgg gcaacacctc tgaggagctg 1920

tgtgtgaggt ggacccagct gggggccttt tatcccttca tgaggaacca caacagcctg 1980

ctgagcctgc ctcaggagcc ctactctttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgaccc tgaggtatgc cctgctgccc cacctgtata ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc caggcccctg ttcctggagt tccccaagga cagcagcacc 2160

tggactgtgg atcatcagct gctgtggggg gaggccctgc tgatcacccc tgtgctgcag 2220

gctggcaagg ctgaggtcac tggctacttc cctctgggca cctggtatga cctgcagact 2280

gtgcctgtgg aggctctggg cagcctgccc cccccccctg ctgctcccag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgctcccc tggacaccat caatgtgcac 2400

ctgaggctg gctacattat ccccctgcag ggcccagggc tgactaccac tgagagcaga 2460

cagcagccca tggctctggc tgtggccctg accaaggggg gggaagctag gggggagctg 2520

ttctgggatg atggggagag cctggaggtg ctggagaggg gggcctatac ccaggtgatc 2580

ttcctggcta ggaacaacac cattgtcaat gagctggtga gggtgacttc tgagggggct 2640

gggctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgctcccca gcaggtgctg 2700

agcaatgggg tgcctgtgag caacttcacc tacagccctg acaccaaggt gctggacatc 2760

tgtgtgtctc tgctgatggg ggagcagttc ctggtgagct ggtgctga 2808

<210> 4

<211> 2808

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 4

atggccttcc tgtggctgct gtcttgctgg gctctgctgg ggaccacctt tggcctgctg 60

gtccccagg agctgtctgg ctcttctcct gtcctggagg agacccaccc tgcccaccag 120

caggggggcta gcaggcctgg ccccagggat gcccaggccc accctggcag gcccaggggct 180

gtgcccaccc agtgtgatgt gcctcccaac agcaggtttg actgtgcccc tgacaaggcc 240

atcacccagg agcagtgtga ggccaggggc tgctgctata tccctgccaa gcagggcctg 300

caggggggctc agatgggcca gccctggtgc ttctttcccc cctcttatcc tagctataag 360

ctggagaacc tgagcagctc tgagatgggg tacactgcca ccctgaccag gaccaccccc 420

actttcttcc ctaaggacat cctgaccctg aggctggatg tgatgatgga gactgagaat 480

aggctgcact ttactatcaa ggaccctgcc aacaggaggt atgaggtgcc tctggagacc 540

ccccatgtgc attctagggc ccccagcccc ctgtactctg tggagttctc tgaggagccc 600

tttggggtga ttgtgaggag acagctggat ggcagggtcc tgctgaacac cactgtggct 660

cccctgtttt ttgctgacca gttcctgcag ctgagcacca gcctgcccag ccagtacatc 720

actgggctgg ctgagcacct gagccccctg atgctgagca ccagctggac caggatcacc 780

ctgtggaaca gggatctggc tcctacccct ggggccaacc tgtatggctc tcaccccttt 840

tacctggccc tggaggatgg gggctctgcc catggggtgt tcctgctgaa cagcaatgct 900

atggatgtgg tgctgcagcc cagccctgcc ctgagctgga ggtctactgg gggcatcctg 960

gatgtgtaca tctttctggg gcctgagccc aagtctgtgg tgcagcagta cctggatgtg 1020

gtgggctatc cttttatgcc cccctattgg ggcctgggct tccacctgtg caggtggggc 1080

tacagcagca ctgccatcac cagacaggtg gtggagaaca tgaccagggc ccacttcccc 1140

ctggatgtgc agtggaatga cctggactac atggacagca ggagggactt cacctttaac 1200

aaggatggct ttagggactt ccctgccatg gtgcaggagc tgcatcaggg gggcaggagg 1260

tacatgatga ttgtggaccc agccatcagc agctctgggc ctgctgggtc ttacaggccc 1320

tatgatgagg gcctgaggag gggggtgttc atcaccaatg agactggcca gcccctgatt 1380

ggcaaggtgt ggcctgggag cactgccttc cctgatttta ccaaccccac tgccctggcc 1440

tggtgggagg atatggtggc tgagtttcat gaccaggtgc cctttgatgg catgtggatt 1500

gacatgaatg agcccagcaa tttcatcagg ggctctgagg atggctgccc caacaatgag 1560

ctggagaatc ctccctatgt gcctggggtg gtggggggca ccctgcaggc tgccaccatc 1620

tgtgcctcta gccaccagtt cctgagcacc cactataacc tgcataacct gtatggcctg 1680

actgaggcca ttgccagcca tagagccctg gtgaaggcca gagggaccag gccctttgtg 1740

atctctagga gcacctttgc tggccatggc aggtatgctg gccactggac tggggatgtg 1800

tggagctctt gggagcagct ggccagctct gtgccagaga tcctgcagtt caacctgctg 1860

ggggtgcctc tggtggggggc tgatgtgtgt ggcttcctgg gcaatacctc tgaagagctg 1920

tgtgtgaggt ggactcagct gggggccttc tatcccttca tgaggaacca caacagcctg 1980

ctgtctctgc cccaggagcc ctacagcttc tctgagcctg ctcagcaggc tatgaggaag 2040

gccctgaccc tgaggtatgc cctgctgccc catctgtaca ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc caggcccctg tttctggagt ttcccaagga cagcagcacc 2160

tggactgtgg accatcagct gctgtggggg gaggctctgc tgattacccc tgtgctgcag 2220

gctggcaagg ctgaggtgac tgggtacttc cccctgggga cttggtatga cctgcagact 2280

gtgcctgtgg aagctctggg cagcctgccc ccaccccctg ctgcccctag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgcccctc tggacaccat caatgtgcac 2400

ctgaggctg gctatatcat ccccctgcag ggccctgggc tgaccaccac tgagagcagg 2460

cagcagccca tggccctggc tgtggccctg actaaggggg gggaggccag gggggagctg 2520

ttctgggatg atggggagag cctggaggtg ctggagagag gggcctacac ccaggtgatc 2580

tttctggcca ggaacaacac cattgtgaat gagctggtga gggtgacttc tgagggggct 2640

ggcctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgcccccca gcaggtgctg 2700

agcaatgggg tgcctgtgtc taacttcacc tacagccctg atactaaggt gctggatatc 2760

tgtgtgagcc tgctgatggg ggagcagttt ctggtgagct ggtgctga 2808

<210> 5

<211> 2808

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 5

atggcctttc tgtggctgct gtcctgctgg gccctgctgg ggaccacctt tggcctgctg 60

gtgcccagg agctgtctgg gagcagccca gtgctggagg agacccaccc tgcccaccag 120

caggggggcca gcaggcctgg ccctagggat gcccaggccc accctggcag gcccaggggct 180

gtgcctaccc agtgtgatgt gccacccaat tctaggtttg actgtgctcc tgacaaggcc 240

atcactcagg agcagtgtga agctaggggg tgctgctaca tcccagccaa gcagggcctg 300

caggggggccc agatgggcca gccctggtgc ttcttccccc ccagctaccc tagctacaag 360

ctggagaatc tgagcagctc tgagatgggc tacactgcta ccctgaccag gaccactcct 420

accttcttcc ccaaggacat cctgactctg aggctggatg tcatgatgga gactgaaaat 480

aggctgcact tcaccatcaa ggaccctgcc aataggaggt atgaggtgcc tctggagacc 540

ccccatgtgc atagcagggc tcccagcccc ctgtattctg tggagttctc tgaggagccc 600

tttggggtca ttgtgaggag acagctggat gggagggtgc tgctgaacac tactgtggct 660

cccctgttct ttgctgacca gttcctgcag ctgtctacca gcctgcccag ccagtacatc 720

actgggctgg ctgagcatct gagccccctg atgctgagca ccagctggac caggatcact 780

ctgtggaaca gggatctggc ccccactcct ggggccaacc tgtatgggag ccatcccttc 840

tacctggccc tggaggatgg gggctctgcc catggggtgt tcctgctgaa cagcaatgcc 900

atggatgtgg tgctgcagcc tagccctgcc ctgagctgga ggagcactgg gggcatcctg 960

gatgtctaca tcttcctggg gcctgagccc aagtctgtgg tgcagcagta tctggatgtg 1020

gtggggtatc ccttcatgcc cccctactgg ggcctgggct ttcacctgtg caggtggggc 1080

tacagcagca ctgccatcac caggcaggtg gtggagaaca tgaccagggc ccacttccct 1140

ctggatgtgc agtggaatga cctggactat atggattcta gggagagactt tacttttaac 1200

aaggatggct tcagggattt ccctgccatg gtgcaggagc tgcaccaggg gggcaggagg 1260

tacatgatga ttgtggaccc tgctattagc agctctggcc ctgctgggtc ttacaggcct 1320

tatgatgagg gcctgaggag gggggtgttc atcaccaatg agactggcca gcccctgatt 1380

ggcaaggtgt ggcctggcag cactgccttc cctgacttca ccaaccccac tgccctggcc 1440

tggtgggagg acatggtggc tgagttccat gaccaggtgc cctttgatgg gatgtggatt 1500

gacatgaatg agccctctaa cttcatcagg gggtctgagg atggctgccc caacaatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ctctgcaggc tgccactatc 1620

tgtgcttctt ctcaccagtt tctgagcacc cactataatc tgcacaacct gtatggcctg 1680

actgaggcca ttgccagcca tagggccctg gtgaaggcca ggggcaccag gccctttgtg 1740

atcagcaggt ctacctttgc tggccatggc aggtatgctg gccactggac tggggatgtg 1800

tggtcttctt ggggagcagct ggccagctct gtgcctgaga tcctgcagtt caacctgctg 1860

ggggtgcctc tggtgggggc tgatgtgtgt ggctttctgg gcaacacctc tgaggagctg 1920

tgtgtgaggt ggacccagct gggggccttt taccccttca tgaggaacca caatagcctg 1980

ctgagcctgc cccaggagcc ttactctttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgactc tgaggtatgc cctgctgccc catctgtata ccctgtttca ccaggcccat 2100

gtggctgggg agactgtggc taggcctctg tttctggagt tccctaagga ctctagcacc 2160

tggactgtgg accaccagct gctgtggggg gaggccctgc tgatcacccc tgtgctgcag 2220

gctggcaagg ctgaggtgac tggctacttc cccctgggca cctggtatga cctgcagact 2280

gtgcctgtgg aggccctggg gagcctgcct cccccccctg ctgcccccag ggagcctgcc 2340

attcattctg agggccagtg ggtgaccctg cctgcccctc tggacaccat caatgtgcac 2400

ctgaggctg ggtacatcat ccccctgcag ggccctggcc tgaccaccac tgagagcagg 2460

cagcagccca tggccctggc tgtggctctg accaaggggg gggaggccag gggggagctg 2520

ttctgggatg atggggagtc tctggaggtg ctggagaggg gggcctacac ccaggtgatc 2580

tttctggcca ggaacaatac tattgtgaat gagctggtga gggtgacctc tgagggggct 2640

ggcctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgcccccca gcaggtcctg 2700

agcaatgggg tgcctgtgag caacttcacc tactctcctg acaccaaggt gctggacatt 2760

tgtgtgtctc tgctgatggg ggagcagttc ctggtgagct ggtgctga 2808

<210> 6

<211> 3016

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 6

atggctttcc tgtggctgct gagctgctgg gctctgctgg gcaccacctt tgggctgctg 60

gtgcctaggg agctgtctgg gtctagccct gtgctggagg agactcaccc tgcccatcag 120

caggggggcta gcaggcctgg ccccagggat gctcaggccc accctggcag gcccaggggct 180

gtgcccaccc agtgtgatgt gccccccaac agcaggtttg actgtgcccc tgacaaggcc 240

attacccagg agcagtgtga ggccaggggc tgctgctaca ttccagctaa gcagggcctg 300

caggggggccc agatgggcca gccctggtgc ttcttccccc ccagctatcc tagctataaa 360

ctggagaacc tgagcagctc tgagatgggc tatactgcca ccctgactag gactactccc 420

accttttttc ctaaggatat cctgaccctg aggctggatg tgatgatgga gactgagaac 480

aggctgcact tcactattaa ggaccctgcc aataggaggt atgaagtgcc tctggagact 540

cctcatgtgc actctagggc ccccagcccc ctgtattctg tggagttctc tgaggagccc 600

tttggggtga ttgtgaggag gcagctggat ggcagggtgc tgctgaacac cactgtggcc 660

cccctgttct ttgctgacca gttcctgcag ctgagcacca gcctgcccag ccagtacatc 720

actgggctgg ctgagcatct gagccctctg atgctgagca cctcttggac caggatcacc 780

ctgtggaata gggatctggc ccccacccct ggggctaatc tgtatggctc tcatcccttt 840

tacctggccc tggaggatgg gggctctgcc catggggtgt ttctgctgaa cagcaatgcc 900

atggatgtgg tgctgcagcc ctctcctgcc ctgagctgga ggagcactgg gggcatcctg 960

gatgtgtaca tcttcctggg ccctgagccc aagtctgtgg tccagcagta tctggatgtg 1020

gtgggctacc cctttatgcc cccctattgg ggcctgggct tccacctgtg caggtggggg 1080

tattcttcta ctgctatcac caggcaggtg gtggagaaca tgaccagggc tcacttcccc 1140

ctggatgtgc agtggaatga cctggactat atggactcta ggagggattt caccttcaac 1200

aaggatggct tcagggactt ccctgctatg gtccaggagc tgcatcaggg gggcaggagg 1260

tacatgatga ttgtggaccc tgccatcagc agctctggcc ctgctggcag ctataggccc 1320

tatgatgagg gcctgaggag gggggtgttt atcactaatg aaactgggca gcccctgatt 1380

ggcaaggtgt ggcctggctc tactgccttc cctgacttca ccaaccccac tgctctggcc 1440

tggtgggagg acatggtggc tgagttccat gaccaggtgc cttttgatgg catgtggatt 1500

gacatgaatg agcccagcaa cttcatcagg ggctctgagg atgggtgccc caataatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ccctgcaggc tgccactatt 1620

tgtgccagct ctcaccagtt cctgagcacc cactacaacc tgcacaatct gtatggcctg 1680

actgaggcca ttgccagcca cagggccctg gtgaaggcca ggggcactag gccctttgtg 1740

atctctagaa gcacctttgc tggccatggg aggtatgctg gccactggac tggggatgtg 1800

tggagctctt gggagcagct ggccagctct gtgcctgaga tcctgcagtt caacctgctg 1860

ggggtgcccc tggtgggggc tgatgtgtgt ggcttcctgg gcaacacctc tgaagagctg 1920

tgtgtgaggt ggacccagct gggggccttc taccctttca tgaggaacca caacagcctg 1980

ctgagcctgc ctcaggagcc ttactctttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgaccc tgaggtatgc tctgctgccc cacctgtaca ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc caggcccctg ttcctggagt ttcctaagga tagcagcacc 2160

tggactgtgg accaccagct gctgtggggg gaggccctgc tgattacccc tgtgctgcag 2220

gctggcaagg ctgaggtgac tggctacttc cccctgggca cttggtatga cctgcagact 2280

gtgcctgtgg aagccctggg cagcctgcct cccccccctg ctgcccccag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgcccccc tggacaccat taatgtgcat 2400

ctgaggctg ggtatattat ccccctgcag gggcctgggc tgactaccac tgagagcagg 2460

cagcagccta tggccctggc tgtggctctg actaaggggg gggaggccag gggggagctg 2520

ttctgggatg atggggagag cctggaggtg ctggagaggg gggcctacac ccaggtgatt 2580

ttcctggcca ggaacaacac cattgtgaat gagctggtga gggtgacctc tgagggggct 2640

ggcctgcagc tgcagaaagt gactgtgctg ggggtggcca ctgcccccca gcaggtgctg 2700

agcaatgggg tgcctgtgag caacttcacc tacagccctg acaccaaggt gctggatatt 2760

tgtgtgagcc tgctgatggg ggagcagttc ctggtgagct ggtgctgact cgagagatct 2820

accggtgaat tcaccgcggg tttaaactgt gccttctagt tgccagccat ctgttgtttg 2880

cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2940

aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3000

gggggctagc tctaga 3016

<210> 7

<211> 3016

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 7

atggctttcc tgtggctgct gtcttgctgg gccctgctgg ggactacctt tggcctgctg 60

gtgcccagg aactgtctgg ctctagccca gtgctggagg agacccaccc tgcccaccag 120

caggggggctt ctaggcctgg ccccagggat gcccaggccc accctggcag gccaagggct 180

gtgcccaccc agtgtgatgt gccccccaac tctagatttg attgtgcccc tgataaggcc 240

atcacccagg agcagtgtga ggctaggggc tgctgctaca tccctgctaa gcagggcctg 300

caggggggctc agatgggcca gccctggtgc ttcttccccc ccagctatcc ctcttacaag 360

ctggagaatc tgagcagctc tgagatgggc tacactgcca ccctgaccag gactactccc 420

accttcttcc ccaaggacat cctgaccctg aggctggatg tgatgatgga gactgagaac 480

aggctgcatt tcaccatcaa ggatcctgcc aacaggaggt atgaggtgcc tctggagacc 540

ccccatgtgc acagcagggc tccttctccc ctgtactctg tggagttctc tgaggaaccc 600

tttggggtga ttgtgaggag gcagctggat ggcagggtcc tgctgaacac cactgtggcc 660

cccctgttct ttgctgatca gttcctgcag ctgtccactt ctctgcctag ccagtacatc 720

actgggctgg ctgagcacct gagccctctg atgctgagca cctcttggac taggatcacc 780

ctgtggaaca gggacctggc ccccacccct ggggccaacc tgtatggcag ccaccccttc 840

tatctggccc tggaggatgg gggctctgcc catggggtgt tcctgctgaa tagcaatgct 900

atggatgtgg tgctgcagcc cagccctgcc ctgtcttgga ggagcactgg gggcatcctg 960

gatgtgtaca ttttcctggg gcctgagccc aagtctgtgg tgcagcagta cctggatgtg 1020

gtgggctacc ccttcatgcc tccctactgg ggcctgggct tccacctgtg caggtggggc 1080

tacagctcta ctgccatcac caggcaggtg gtggagaata tgaccagggc ccacttcccc 1140

ctggatgtgc agtggaatga cctggactac atggactcta ggagggactt caccttcaat 1200

aaggatggct tcagagactt ccctgccatg gtgcaggagc tgcatcaggg gggcaggagg 1260

tacatgatga ttgtggaccc tgccatcagc tcttctggcc ctgctggctc ttacaggccc 1320

tatgatgagg gcctgaggag gggggtgttc atcaccaatg agactgggca gcccctgatt 1380

gggaaggtgt ggcctggctc tactgccttc cctgacttca ccaatcctac tgccctggcc 1440

tggtgggagg acatggtggc tgagttccat gaccaggtgc cctttgatgg catgtggatt 1500

gacatgaatg agccctctaa tttcatcagg ggctctgagg atggctgccc caacaatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ccctgcaggc tgccaccatc 1620

tgtgctagct ctcaccagtt cctgagcacc cactacaatc tgcataacct gtatggcctg 1680

actgaggcca ttgccagcca cagggccctg gtgaaggcta ggggcaccag gccctttgtg 1740

atttctagga gcacttttgc tggccatggc aggtatgctg ggcactggac tggggatgtg 1800

tggtctagct gggagcagct ggcttcttct gtgcctgaga tcctgcagtt caacctgctg 1860

ggggtgcctc tggtgggggc tgatgtgtgt gggttcctgg gcaacacttc tgaggagctg 1920

tgtgtgaggt ggacccagct gggggccttc taccctttca tgaggaacca caacagcctg 1980

ctgagcctgc cccaggagcc ctacagcttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgaccc tgaggtatgc cctgctgccc cacctgtaca ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc taggcctctg ttcctggagt tccccaagga ctctagcacc 2160

tggactgtgg accaccagct gctgtggggg gaggccctgc tgatcactcc tgtgctgcag 2220

gctgggaagg ctgaggtgac tggctatttc cccctgggca cctggtatga cctgcagact 2280

gtgcctgtgg aggccctggg gagcctgccc cccccccctg ctgcccccag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgcccctc tggataccat caatgtgcac 2400

ctgaggctg gctacatcat tcccctgcag ggccctggcc tgaccactac tgagtctagg 2460

cagcagccca tggccctggc tgtggccctg accaaggggg gggaggctag gggggagctg 2520

ttttgggatg atggggagag cctggaggtg ctggagaggg gggcctacac tcaggtgatc 2580

ttcctggcca ggaacaatac cattgtgaat gagctggtga gggtgacctc tgagggggct 2640

ggcctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgcccccca gcaggtgctg 2700

agcaatgggg tgcctgtgag caacttcacc tatagccctg ataccaaggt gctggatatt 2760

tgtgtgagcc tgctgatggg ggagcagttc ctggtgagct ggtgctgact cgagagatct 2820

accggtgaat tcaccgcggg tttaaactgt gccttctagt tgccagccat ctgttgtttg 2880

cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2940

aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3000

gggggctagc tctaga 3016

<210> 8

<211> 3016

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 8

atggctttcc tgtggctgct gtcttgttgg gctctgctgg gcaccacctt tggcctgctg 60

gtgcccaggg agctgtctgg cagcagccct gtgctggagg agacccaccc tgctcatcag 120

caggggggcta gcaggcctgg ccccagggat gcccaggctc accctgggag acccagggct 180

gtgcccactc agtgtgatgt gccccccaac agcaggtttg actgtgctcc tgacaaggct 240

atcacccagg agcagtgtga ggccagggg tgctgctaca ttcctgctaa gcagggcctg 300

caggggggccc agatgggcca gccctggtgc ttcttccccc cctcttatcc cagctataag 360

ctggagaacc tgagcagctc tgagatgggc tacactgcca ccctgaccag gaccactccc 420

accttctttc ccaaggatat tctgactctg aggctggatg tgatgatgga gactgagaac 480

aggctgcact tcactatcaa ggaccctgcc aataggaggt atgaggtgcc cctggagact 540

cctcatgtgc atagcagggc cccttctcct ctgtattctg tggagttctc tgaggagccc 600

tttggggtga ttgtgaggag gcagctggat ggcagggtgc tgctgaacac cactgtggcc 660

cccctgttct ttgctgacca gttcctgcag ctgagcactt ctctgcccag ccagtacatt 720

actgggctgg ctgagcatct gagccccctg atgctgagca cctcttggac caggatcacc 780

ctgtggaaca gggacctggc ccccactcct ggggctaacc tgtatggctc tcaccccttt 840

tacctggccc tggaggatgg gggctctgcc catggggtgt ttctgctgaa cagcaatgct 900

atggatgtgg tgctgcagcc ctctccagcc ctgtcttgga ggagcactgg gggcattctg 960

gatgtgtaca ttttcctggg gcctgaaccc aagtctgtgg tgcagcagta cctggatgtg 1020

gtgggctacc ccttcatgcc cccctattgg gggctggggt ttcacctgtg caggtggggc 1080

tacagcagca ctgccatcac caggcaggtg gtggagaaca tgaccagggc ccatttcccc 1140

ctggatgtgc agtggaatga cctggactac atggatagca ggagggattt caccttcaac 1200

aaggatggct tcagggactt tcctgccatg gtgcaggagc tgcaccaggg gggcaggagg 1260

tatatgatga ttgtggaccc tgctatcagc agctctggcc ctgctggctc ttacaggccc 1320

tatgatgagg gcctgaggag gggggtgttt atcactaatg aaactggcca gcctctgatt 1380

ggcaaggtct ggcctggctc tactgccttc cctgatttta ctaaccccac tgccctggcc 1440

tggtgggagg acatggtggc tgagttccat gatcaggtgc cttttgatgg catgtggatt 1500

gatatgaatg aaccaagcaa cttcatcaga ggctctgagg atggctgccc caacaatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ctctgcaggc tgccaccatt 1620

tgtgctagca gccaccagtt cctgagcacc cactacaatc tgcacaacct gtatggcctg 1680

actgaagcca ttgccagcca tagggccctg gtgaaggcca ggggcactag gccttttgtg 1740

atcagcagga gcacttttgc tggccatggc aggtatgctg gccactggac tggggatgtg 1800

tggagcagct gggagcagct ggccagctct gtgcctgaga ttctgcagtt taacctgctg 1860

ggggtgcccc tggtgggggc tgatgtgtgt ggcttcctgg gcaacacctc tgaggagctg 1920

tgtgtgaggt ggacccagct gggggccttt tatcccttca tgaggaacca caacagcctg 1980

ctgagcctgc ctcaggagcc ctactctttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgaccc tgaggtatgc cctgctgccc cacctgtata ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc caggcccctg ttcctggagt tccccaagga cagcagcacc 2160

tggactgtgg atcatcagct gctgtggggg gaggccctgc tgatcacccc tgtgctgcag 2220

gctggcaagg ctgaggtcac tggctacttc cctctgggca cctggtatga cctgcagact 2280

gtgcctgtgg aggctctggg cagcctgccc cccccccctg ctgctcccag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgctcccc tggacaccat caatgtgcac 2400

ctgaggctg gctacattat ccccctgcag ggcccagggc tgactaccac tgagagcaga 2460

cagcagccca tggctctggc tgtggccctg accaaggggg gggaagctag gggggagctg 2520

ttctgggatg atggggagag cctggaggtg ctggagaggg gggcctatac ccaggtgatc 2580

ttcctggcta ggaacaacac cattgtcaat gagctggtga gggtgacttc tgagggggct 2640

gggctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgctcccca gcaggtgctg 2700

agcaatgggg tgcctgtgag caacttcacc tacagccctg acaccaaggt gctggacatc 2760

tgtgtgtctc tgctgatggg ggagcagttc ctggtgagct ggtgctgact cgagagatct 2820

accggtgaat tcaccgcggg tttaaactgt gccttctagt tgccagccat ctgttgtttg 2880

cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2940

aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3000

gggggctagc tctaga 3016

<210> 9

<211> 3016

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 9

atggccttcc tgtggctgct gtcttgctgg gctctgctgg ggaccacctt tggcctgctg 60

gtccccagg agctgtctgg ctcttctcct gtcctggagg agacccaccc tgcccaccag 120

caggggggcta gcaggcctgg ccccagggat gcccaggccc accctggcag gcccaggggct 180

gtgcccaccc agtgtgatgt gcctcccaac agcaggtttg actgtgcccc tgacaaggcc 240

atcacccagg agcagtgtga ggccaggggc tgctgctata tccctgccaa gcagggcctg 300

caggggggctc agatgggcca gccctggtgc ttctttcccc cctcttatcc tagctataag 360

ctggagaacc tgagcagctc tgagatgggg tacactgcca ccctgaccag gaccaccccc 420

actttcttcc ctaaggacat cctgaccctg aggctggatg tgatgatgga gactgagaat 480

aggctgcact ttactatcaa ggaccctgcc aacaggaggt atgaggtgcc tctggagacc 540

ccccatgtgc attctagggc ccccagcccc ctgtactctg tggagttctc tgaggagccc 600

tttggggtga ttgtgaggag acagctggat ggcagggtcc tgctgaacac cactgtggct 660

cccctgtttt ttgctgacca gttcctgcag ctgagcacca gcctgcccag ccagtacatc 720

actgggctgg ctgagcacct gagccccctg atgctgagca ccagctggac caggatcacc 780

ctgtggaaca gggatctggc tcctacccct ggggccaacc tgtatggctc tcaccccttt 840

tacctggccc tggaggatgg gggctctgcc catggggtgt tcctgctgaa cagcaatgct 900

atggatgtgg tgctgcagcc cagccctgcc ctgagctgga ggtctactgg gggcatcctg 960

gatgtgtaca tctttctggg gcctgagccc aagtctgtgg tgcagcagta cctggatgtg 1020

gtgggctatc cttttatgcc cccctattgg ggcctgggct tccacctgtg caggtggggc 1080

tacagcagca ctgccatcac cagacaggtg gtggagaaca tgaccagggc ccacttcccc 1140

ctggatgtgc agtggaatga cctggactac atggacagca ggagggactt cacctttaac 1200

aaggatggct ttagggactt ccctgccatg gtgcaggagc tgcatcaggg gggcaggagg 1260

tacatgatga ttgtggaccc agccatcagc agctctgggc ctgctgggtc ttacaggccc 1320

tatgatgagg gcctgaggag gggggtgttc atcaccaatg agactggcca gcccctgatt 1380

ggcaaggtgt ggcctgggag cactgccttc cctgatttta ccaaccccac tgccctggcc 1440

tggtgggagg atatggtggc tgagtttcat gaccaggtgc cctttgatgg catgtggatt 1500

gacatgaatg agcccagcaa tttcatcagg ggctctgagg atggctgccc caacaatgag 1560

ctggagaatc ctccctatgt gcctggggtg gtggggggca ccctgcaggc tgccaccatc 1620

tgtgcctcta gccaccagtt cctgagcacc cactataacc tgcataacct gtatggcctg 1680

actgaggcca ttgccagcca tagagccctg gtgaaggcca gagggaccag gccctttgtg 1740

atctctagga gcacctttgc tggccatggc aggtatgctg gccactggac tggggatgtg 1800

tggagctctt gggagcagct ggccagctct gtgccagaga tcctgcagtt caacctgctg 1860

ggggtgcctc tggtggggggc tgatgtgtgt ggcttcctgg gcaatacctc tgaagagctg 1920

tgtgtgaggt ggactcagct gggggccttc tatcccttca tgaggaacca caacagcctg 1980

ctgtctctgc cccaggagcc ctacagcttc tctgagcctg ctcagcaggc tatgaggaag 2040

gccctgaccc tgaggtatgc cctgctgccc catctgtaca ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc caggcccctg tttctggagt ttcccaagga cagcagcacc 2160

tggactgtgg accatcagct gctgtggggg gaggctctgc tgattacccc tgtgctgcag 2220

gctggcaagg ctgaggtgac tgggtacttc cccctgggga cttggtatga cctgcagact 2280

gtgcctgtgg aagctctggg cagcctgccc ccaccccctg ctgcccctag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgcccctc tggacaccat caatgtgcac 2400

ctgaggctg gctatatcat ccccctgcag ggccctgggc tgaccaccac tgagagcagg 2460

cagcagccca tggccctggc tgtggccctg actaaggggg gggaggccag gggggagctg 2520

ttctgggatg atggggagag cctggaggtg ctggagagag gggcctacac ccaggtgatc 2580

tttctggcca ggaacaacac cattgtgaat gagctggtga gggtgacttc tgagggggct 2640

ggcctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgcccccca gcaggtgctg 2700

agcaatgggg tgcctgtgtc taacttcacc tacagccctg atactaaggt gctggatatc 2760

tgtgtgagcc tgctgatggg ggagcagttt ctggtgagct ggtgctgact cgagagatct 2820

accggtgaat tcaccgcggg tttaaactgt gccttctagt tgccagccat ctgttgtttg 2880

cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2940

aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3000

gggggctagc tctaga 3016

<210> 10

<211> 3016

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 10

atggcctttc tgtggctgct gtcctgctgg gccctgctgg ggaccacctt tggcctgctg 60

gtgcccagg agctgtctgg gagcagccca gtgctggagg agacccaccc tgcccaccag 120

caggggggcca gcaggcctgg ccctagggat gcccaggccc accctggcag gcccaggggct 180

gtgcctaccc agtgtgatgt gccacccaat tctaggtttg actgtgctcc tgacaaggcc 240

atcactcagg agcagtgtga agctaggggg tgctgctaca tcccagccaa gcagggcctg 300

caggggggccc agatgggcca gccctggtgc ttcttccccc ccagctaccc tagctacaag 360

ctggagaatc tgagcagctc tgagatgggc tacactgcta ccctgaccag gaccactcct 420

accttcttcc ccaaggacat cctgactctg aggctggatg tcatgatgga gactgaaaat 480

aggctgcact tcaccatcaa ggaccctgcc aataggaggt atgaggtgcc tctggagacc 540

ccccatgtgc atagcagggc tcccagcccc ctgtattctg tggagttctc tgaggagccc 600

tttggggtca ttgtgaggag acagctggat gggagggtgc tgctgaacac tactgtggct 660

cccctgttct ttgctgacca gttcctgcag ctgtctacca gcctgcccag ccagtacatc 720

actgggctgg ctgagcatct gagccccctg atgctgagca ccagctggac caggatcact 780

ctgtggaaca gggatctggc ccccactcct ggggccaacc tgtatgggag ccatcccttc 840

tacctggccc tggaggatgg gggctctgcc catggggtgt tcctgctgaa cagcaatgcc 900

atggatgtgg tgctgcagcc tagccctgcc ctgagctgga ggagcactgg gggcatcctg 960

gatgtctaca tcttcctggg gcctgagccc aagtctgtgg tgcagcagta tctggatgtg 1020

gtggggtatc ccttcatgcc cccctactgg ggcctgggct ttcacctgtg caggtggggc 1080

tacagcagca ctgccatcac caggcaggtg gtggagaaca tgaccagggc ccacttccct 1140

ctggatgtgc agtggaatga cctggactat atggattcta gggagagactt tacttttaac 1200

aaggatggct tcagggattt ccctgccatg gtgcaggagc tgcaccaggg gggcaggagg 1260

tacatgatga ttgtggaccc tgctattagc agctctggcc ctgctgggtc ttacaggcct 1320

tatgatgagg gcctgaggag gggggtgttc atcaccaatg agactggcca gcccctgatt 1380

ggcaaggtgt ggcctggcag cactgccttc cctgacttca ccaaccccac tgccctggcc 1440

tggtgggagg acatggtggc tgagttccat gaccaggtgc cctttgatgg gatgtggatt 1500

gacatgaatg agccctctaa cttcatcagg gggtctgagg atggctgccc caacaatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ctctgcaggc tgccactatc 1620

tgtgcttctt ctcaccagtt tctgagcacc cactataatc tgcacaacct gtatggcctg 1680

actgaggcca ttgccagcca tagggccctg gtgaaggcca ggggcaccag gccctttgtg 1740

atcagcaggt ctacctttgc tggccatggc aggtatgctg gccactggac tggggatgtg 1800

tggtcttctt ggggagcagct ggccagctct gtgcctgaga tcctgcagtt caacctgctg 1860

ggggtgcctc tggtgggggc tgatgtgtgt ggctttctgg gcaacacctc tgaggagctg 1920

tgtgtgaggt ggacccagct gggggccttt taccccttca tgaggaacca caatagcctg 1980

ctgagcctgc cccaggagcc ttactctttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgactc tgaggtatgc cctgctgccc catctgtata ccctgtttca ccaggcccat 2100

gtggctgggg agactgtggc taggcctctg tttctggagt tccctaagga ctctagcacc 2160

tggactgtgg accaccagct gctgtggggg gaggccctgc tgatcacccc tgtgctgcag 2220

gctggcaagg ctgaggtgac tggctacttc cccctgggca cctggtatga cctgcagact 2280

gtgcctgtgg aggccctggg gagcctgcct cccccccctg ctgcccccag ggagcctgcc 2340

attcattctg agggccagtg ggtgaccctg cctgcccctc tggacaccat caatgtgcac 2400

ctgaggctg ggtacatcat ccccctgcag ggccctggcc tgaccaccac tgagagcagg 2460

cagcagccca tggccctggc tgtggctctg accaaggggg gggaggccag gggggagctg 2520

ttctgggatg atggggagtc tctggaggtg ctggagaggg gggcctacac ccaggtgatc 2580

tttctggcca ggaacaatac tattgtgaat gagctggtga gggtgacctc tgagggggct 2640

ggcctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgcccccca gcaggtcctg 2700

agcaatgggg tgcctgtgag caacttcacc tactctcctg acaccaaggt gctggacatt 2760

tgtgtgtctc tgctgatggg ggagcagttc ctggtgagct ggtgctgact cgagagatct 2820

accggtgaat tcaccgcggg tttaaactgt gccttctagt tgccagccat ctgttgtttg 2880

cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2940

aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3000

gggggctagc tctaga 3016

<210> 11

<211> 3076

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 11

atggctttcc tgtggctgct gagctgctgg gctctgctgg gcaccacctt tgggctgctg 60

gtgcctaggg agctgtctgg gtctagccct gtgctggagg agactcaccc tgcccatcag 120

caggggggcta gcaggcctgg ccccagggat gctcaggccc accctggcag gcccaggggct 180

gtgcccaccc agtgtgatgt gccccccaac agcaggtttg actgtgcccc tgacaaggcc 240

attacccagg agcagtgtga ggccaggggc tgctgctaca ttccagctaa gcagggcctg 300

caggggggccc agatgggcca gccctggtgc ttcttccccc ccagctatcc tagctataaa 360

ctggagaacc tgagcagctc tgagatgggc tatactgcca ccctgactag gactactccc 420

accttttttc ctaaggatat cctgaccctg aggctggatg tgatgatgga gactgagaac 480

aggctgcact tcactattaa ggaccctgcc aataggaggt atgaagtgcc tctggagact 540

cctcatgtgc actctagggc ccccagcccc ctgtattctg tggagttctc tgaggagccc 600

tttggggtga ttgtgaggag gcagctggat ggcagggtgc tgctgaacac cactgtggcc 660

cccctgttct ttgctgacca gttcctgcag ctgagcacca gcctgcccag ccagtacatc 720

actgggctgg ctgagcatct gagccctctg atgctgagca cctcttggac caggatcacc 780

ctgtggaata gggatctggc ccccacccct ggggctaatc tgtatggctc tcatcccttt 840

tacctggccc tggaggatgg gggctctgcc catggggtgt ttctgctgaa cagcaatgcc 900

atggatgtgg tgctgcagcc ctctcctgcc ctgagctgga ggagcactgg gggcatcctg 960

gatgtgtaca tcttcctggg ccctgagccc aagtctgtgg tccagcagta tctggatgtg 1020

gtgggctacc cctttatgcc cccctattgg ggcctgggct tccacctgtg caggtggggg 1080

tattcttcta ctgctatcac caggcaggtg gtggagaaca tgaccagggc tcacttcccc 1140

ctggatgtgc agtggaatga cctggactat atggactcta ggagggattt caccttcaac 1200

aaggatggct tcagggactt ccctgctatg gtccaggagc tgcatcaggg gggcaggagg 1260

tacatgatga ttgtggaccc tgccatcagc agctctggcc ctgctggcag ctataggccc 1320

tatgatgagg gcctgaggag gggggtgttt atcactaatg aaactgggca gcccctgatt 1380

ggcaaggtgt ggcctggctc tactgccttc cctgacttca ccaaccccac tgctctggcc 1440

tggtgggagg acatggtggc tgagttccat gaccaggtgc cttttgatgg catgtggatt 1500

gacatgaatg agcccagcaa cttcatcagg ggctctgagg atgggtgccc caataatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ccctgcaggc tgccactatt 1620

tgtgccagct ctcaccagtt cctgagcacc cactacaacc tgcacaatct gtatggcctg 1680

actgaggcca ttgccagcca cagggccctg gtgaaggcca ggggcactag gccctttgtg 1740

atctctagaa gcacctttgc tggccatggg aggtatgctg gccactggac tggggatgtg 1800

tggagctctt gggagcagct ggccagctct gtgcctgaga tcctgcagtt caacctgctg 1860

ggggtgcccc tggtgggggc tgatgtgtgt ggcttcctgg gcaacacctc tgaagagctg 1920

tgtgtgaggt ggacccagct gggggccttc taccctttca tgaggaacca caacagcctg 1980

ctgagcctgc ctcaggagcc ttactctttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgaccc tgaggtatgc tctgctgccc cacctgtaca ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc caggcccctg ttcctggagt ttcctaagga tagcagcacc 2160

tggactgtgg accaccagct gctgtggggg gaggccctgc tgattacccc tgtgctgcag 2220

gctggcaagg ctgaggtgac tggctacttc cccctgggca cttggtatga cctgcagact 2280

gtgcctgtgg aagccctggg cagcctgcct cccccccctg ctgcccccag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgcccccc tggacaccat taatgtgcat 2400

ctgaggctg ggtatattat ccccctgcag gggcctgggc tgactaccac tgagagcagg 2460

cagcagccta tggccctggc tgtggctctg actaaggggg gggaggccag gggggagctg 2520

ttctgggatg atggggagag cctggaggtg ctggagaggg gggcctacac ccaggtgatt 2580

ttcctggcca ggaacaacac cattgtgaat gagctggtga gggtgacctc tgagggggct 2640

ggcctgcagc tgcagaaagt gactgtgctg ggggtggcca ctgcccccca gcaggtgctg 2700

agcaatgggg tgcctgtgag caacttcacc tacagccctg acaccaaggt gctggatatt 2760

tgtgtgagcc tgctgatggg ggagcagttc ctggtgagct ggtgctgaag atctagagct 2820

gaattcctgc agccagggg atcagcctct actgtgcctt ctagttgcca gccatctgtt 2880

gtttgcccct cccccttgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 2940

taataaaatg aggaaattgc atcacattgt ctgagtaggt gtcattctat tctggggggt 3000

ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 3060

gcagtgggct ctatgg 3076

<210> 12

<211> 3076

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 12

atggctttcc tgtggctgct gtcttgctgg gccctgctgg ggactacctt tggcctgctg 60

gtgcccagg aactgtctgg ctctagccca gtgctggagg agacccaccc tgcccaccag 120

caggggggctt ctaggcctgg ccccagggat gcccaggccc accctggcag gccaagggct 180

gtgcccaccc agtgtgatgt gccccccaac tctagatttg attgtgcccc tgataaggcc 240

atcacccagg agcagtgtga ggctaggggc tgctgctaca tccctgctaa gcagggcctg 300

caggggggctc agatgggcca gccctggtgc ttcttccccc ccagctatcc ctcttacaag 360

ctggagaatc tgagcagctc tgagatgggc tacactgcca ccctgaccag gactactccc 420

accttcttcc ccaaggacat cctgaccctg aggctggatg tgatgatgga gactgagaac 480

aggctgcatt tcaccatcaa ggatcctgcc aacaggaggt atgaggtgcc tctggagacc 540

ccccatgtgc acagcagggc tccttctccc ctgtactctg tggagttctc tgaggaaccc 600

tttggggtga ttgtgaggag gcagctggat ggcagggtcc tgctgaacac cactgtggcc 660

cccctgttct ttgctgatca gttcctgcag ctgtccactt ctctgcctag ccagtacatc 720

actgggctgg ctgagcacct gagccctctg atgctgagca cctcttggac taggatcacc 780

ctgtggaaca gggacctggc ccccacccct ggggccaacc tgtatggcag ccaccccttc 840

tatctggccc tggaggatgg gggctctgcc catggggtgt tcctgctgaa tagcaatgct 900

atggatgtgg tgctgcagcc cagccctgcc ctgtcttgga ggagcactgg gggcatcctg 960

gatgtgtaca ttttcctggg gcctgagccc aagtctgtgg tgcagcagta cctggatgtg 1020

gtgggctacc ccttcatgcc tccctactgg ggcctgggct tccacctgtg caggtggggc 1080

tacagctcta ctgccatcac caggcaggtg gtggagaata tgaccagggc ccacttcccc 1140

ctggatgtgc agtggaatga cctggactac atggactcta ggagggactt caccttcaat 1200

aaggatggct tcagagactt ccctgccatg gtgcaggagc tgcatcaggg gggcaggagg 1260

tacatgatga ttgtggaccc tgccatcagc tcttctggcc ctgctggctc ttacaggccc 1320

tatgatgagg gcctgaggag gggggtgttc atcaccaatg agactgggca gcccctgatt 1380

gggaaggtgt ggcctggctc tactgccttc cctgacttca ccaatcctac tgccctggcc 1440

tggtgggagg acatggtggc tgagttccat gaccaggtgc cctttgatgg catgtggatt 1500

gacatgaatg agccctctaa tttcatcagg ggctctgagg atggctgccc caacaatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ccctgcaggc tgccaccatc 1620

tgtgctagct ctcaccagtt cctgagcacc cactacaatc tgcataacct gtatggcctg 1680

actgaggcca ttgccagcca cagggccctg gtgaaggcta ggggcaccag gccctttgtg 1740

atttctagga gcacttttgc tggccatggc aggtatgctg ggcactggac tggggatgtg 1800

tggtctagct gggagcagct ggcttcttct gtgcctgaga tcctgcagtt caacctgctg 1860

ggggtgcctc tggtgggggc tgatgtgtgt gggttcctgg gcaacacttc tgaggagctg 1920

tgtgtgaggt ggacccagct gggggccttc taccctttca tgaggaacca caacagcctg 1980

ctgagcctgc cccaggagcc ctacagcttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgaccc tgaggtatgc cctgctgccc cacctgtaca ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc taggcctctg ttcctggagt tccccaagga ctctagcacc 2160

tggactgtgg accaccagct gctgtggggg gaggccctgc tgatcactcc tgtgctgcag 2220

gctgggaagg ctgaggtgac tggctatttc cccctgggca cctggtatga cctgcagact 2280

gtgcctgtgg aggccctggg gagcctgccc cccccccctg ctgcccccag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgcccctc tggataccat caatgtgcac 2400

ctgaggctg gctacatcat tcccctgcag ggccctggcc tgaccactac tgagtctagg 2460

cagcagccca tggccctggc tgtggccctg accaaggggg gggaggctag gggggagctg 2520

ttttgggatg atggggagag cctggaggtg ctggagaggg gggcctacac tcaggtgatc 2580

ttcctggcca ggaacaatac cattgtgaat gagctggtga gggtgacctc tgagggggct 2640

ggcctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgcccccca gcaggtgctg 2700

agcaatgggg tgcctgtgag caacttcacc tatagccctg ataccaaggt gctggatatt 2760

tgtgtgagcc tgctgatggg ggagcagttc ctggtgagct ggtgctgaag atctagagct 2820

gaattcctgc agccagggg atcagcctct actgtgcctt ctagttgcca gccatctgtt 2880

gtttgcccct cccccttgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 2940

taataaaatg aggaaattgc atcacattgt ctgagtaggt gtcattctat tctggggggt 3000

ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 3060

gcagtgggct ctatgg 3076

<210> 13

<211> 3076

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 13

atggctttcc tgtggctgct gtcttgttgg gctctgctgg gcaccacctt tggcctgctg 60

gtgcccaggg agctgtctgg cagcagccct gtgctggagg agacccaccc tgctcatcag 120

caggggggcta gcaggcctgg ccccagggat gcccaggctc accctgggag acccagggct 180

gtgcccactc agtgtgatgt gccccccaac agcaggtttg actgtgctcc tgacaaggct 240

atcacccagg agcagtgtga ggccagggg tgctgctaca ttcctgctaa gcagggcctg 300

caggggggccc agatgggcca gccctggtgc ttcttccccc cctcttatcc cagctataag 360

ctggagaacc tgagcagctc tgagatgggc tacactgcca ccctgaccag gaccactccc 420

accttctttc ccaaggatat tctgactctg aggctggatg tgatgatgga gactgagaac 480

aggctgcact tcactatcaa ggaccctgcc aataggaggt atgaggtgcc cctggagact 540

cctcatgtgc atagcagggc cccttctcct ctgtattctg tggagttctc tgaggagccc 600

tttggggtga ttgtgaggag gcagctggat ggcagggtgc tgctgaacac cactgtggcc 660

cccctgttct ttgctgacca gttcctgcag ctgagcactt ctctgcccag ccagtacatt 720

actgggctgg ctgagcatct gagccccctg atgctgagca cctcttggac caggatcacc 780

ctgtggaaca gggacctggc ccccactcct ggggctaacc tgtatggctc tcaccccttt 840

tacctggccc tggaggatgg gggctctgcc catggggtgt ttctgctgaa cagcaatgct 900

atggatgtgg tgctgcagcc ctctccagcc ctgtcttgga ggagcactgg gggcattctg 960

gatgtgtaca ttttcctggg gcctgaaccc aagtctgtgg tgcagcagta cctggatgtg 1020

gtgggctacc ccttcatgcc cccctattgg gggctggggt ttcacctgtg caggtggggc 1080

tacagcagca ctgccatcac caggcaggtg gtggagaaca tgaccagggc ccatttcccc 1140

ctggatgtgc agtggaatga cctggactac atggatagca ggagggattt caccttcaac 1200

aaggatggct tcagggactt tcctgccatg gtgcaggagc tgcaccaggg gggcaggagg 1260

tatatgatga ttgtggaccc tgctatcagc agctctggcc ctgctggctc ttacaggccc 1320

tatgatgagg gcctgaggag gggggtgttt atcactaatg aaactggcca gcctctgatt 1380

ggcaaggtct ggcctggctc tactgccttc cctgatttta ctaaccccac tgccctggcc 1440

tggtgggagg acatggtggc tgagttccat gatcaggtgc cttttgatgg catgtggatt 1500

gatatgaatg aaccaagcaa cttcatcaga ggctctgagg atggctgccc caacaatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ctctgcaggc tgccaccatt 1620

tgtgctagca gccaccagtt cctgagcacc cactacaatc tgcacaacct gtatggcctg 1680

actgaagcca ttgccagcca tagggccctg gtgaaggcca ggggcactag gccttttgtg 1740

atcagcagga gcacttttgc tggccatggc aggtatgctg gccactggac tggggatgtg 1800

tggagcagct gggagcagct ggccagctct gtgcctgaga ttctgcagtt taacctgctg 1860

ggggtgcccc tggtgggggc tgatgtgtgt ggcttcctgg gcaacacctc tgaggagctg 1920

tgtgtgaggt ggacccagct gggggccttt tatcccttca tgaggaacca caacagcctg 1980

ctgagcctgc ctcaggagcc ctactctttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgaccc tgaggtatgc cctgctgccc cacctgtata ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc caggcccctg ttcctggagt tccccaagga cagcagcacc 2160

tggactgtgg atcatcagct gctgtggggg gaggccctgc tgatcacccc tgtgctgcag 2220

gctggcaagg ctgaggtcac tggctacttc cctctgggca cctggtatga cctgcagact 2280

gtgcctgtgg aggctctggg cagcctgccc cccccccctg ctgctcccag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgctcccc tggacaccat caatgtgcac 2400

ctgaggctg gctacattat ccccctgcag ggcccagggc tgactaccac tgagagcaga 2460

cagcagccca tggctctggc tgtggccctg accaaggggg gggaagctag gggggagctg 2520

ttctgggatg atggggagag cctggaggtg ctggagaggg gggcctatac ccaggtgatc 2580

ttcctggcta ggaacaacac cattgtcaat gagctggtga gggtgacttc tgagggggct 2640

gggctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgctcccca gcaggtgctg 2700

agcaatgggg tgcctgtgag caacttcacc tacagccctg acaccaaggt gctggacatc 2760

tgtgtgtctc tgctgatggg ggagcagttc ctggtgagct ggtgctgaag atctagagct 2820

gaattcctgc agccagggg atcagcctct actgtgcctt ctagttgcca gccatctgtt 2880

gtttgcccct cccccttgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 2940

taataaaatg aggaaattgc atcacattgt ctgagtaggt gtcattctat tctggggggt 3000

ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 3060

gcagtgggct ctatgg 3076

<210> 14

<211> 3076

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 14

atggccttcc tgtggctgct gtcttgctgg gctctgctgg ggaccacctt tggcctgctg 60

gtccccagg agctgtctgg ctcttctcct gtcctggagg agacccaccc tgcccaccag 120

caggggggcta gcaggcctgg ccccagggat gcccaggccc accctggcag gcccaggggct 180

gtgcccaccc agtgtgatgt gcctcccaac agcaggtttg actgtgcccc tgacaaggcc 240

atcacccagg agcagtgtga ggccaggggc tgctgctata tccctgccaa gcagggcctg 300

caggggggctc agatgggcca gccctggtgc ttctttcccc cctcttatcc tagctataag 360

ctggagaacc tgagcagctc tgagatgggg tacactgcca ccctgaccag gaccaccccc 420

actttcttcc ctaaggacat cctgaccctg aggctggatg tgatgatgga gactgagaat 480

aggctgcact ttactatcaa ggaccctgcc aacaggaggt atgaggtgcc tctggagacc 540

ccccatgtgc attctagggc ccccagcccc ctgtactctg tggagttctc tgaggagccc 600

tttggggtga ttgtgaggag acagctggat ggcagggtcc tgctgaacac cactgtggct 660

cccctgtttt ttgctgacca gttcctgcag ctgagcacca gcctgcccag ccagtacatc 720

actgggctgg ctgagcacct gagccccctg atgctgagca ccagctggac caggatcacc 780

ctgtggaaca gggatctggc tcctacccct ggggccaacc tgtatggctc tcaccccttt 840

tacctggccc tggaggatgg gggctctgcc catggggtgt tcctgctgaa cagcaatgct 900

atggatgtgg tgctgcagcc cagccctgcc ctgagctgga ggtctactgg gggcatcctg 960

gatgtgtaca tctttctggg gcctgagccc aagtctgtgg tgcagcagta cctggatgtg 1020

gtgggctatc cttttatgcc cccctattgg ggcctgggct tccacctgtg caggtggggc 1080

tacagcagca ctgccatcac cagacaggtg gtggagaaca tgaccagggc ccacttcccc 1140

ctggatgtgc agtggaatga cctggactac atggacagca ggagggactt cacctttaac 1200

aaggatggct ttagggactt ccctgccatg gtgcaggagc tgcatcaggg gggcaggagg 1260

tacatgatga ttgtggaccc agccatcagc agctctgggc ctgctgggtc ttacaggccc 1320

tatgatgagg gcctgaggag gggggtgttc atcaccaatg agactggcca gcccctgatt 1380

ggcaaggtgt ggcctgggag cactgccttc cctgatttta ccaaccccac tgccctggcc 1440

tggtgggagg atatggtggc tgagtttcat gaccaggtgc cctttgatgg catgtggatt 1500

gacatgaatg agcccagcaa tttcatcagg ggctctgagg atggctgccc caacaatgag 1560

ctggagaatc ctccctatgt gcctggggtg gtggggggca ccctgcaggc tgccaccatc 1620

tgtgcctcta gccaccagtt cctgagcacc cactataacc tgcataacct gtatggcctg 1680

actgaggcca ttgccagcca tagagccctg gtgaaggcca gagggaccag gccctttgtg 1740

atctctagga gcacctttgc tggccatggc aggtatgctg gccactggac tggggatgtg 1800

tggagctctt gggagcagct ggccagctct gtgccagaga tcctgcagtt caacctgctg 1860

ggggtgcctc tggtggggggc tgatgtgtgt ggcttcctgg gcaatacctc tgaagagctg 1920

tgtgtgaggt ggactcagct gggggccttc tatcccttca tgaggaacca caacagcctg 1980

ctgtctctgc cccaggagcc ctacagcttc tctgagcctg ctcagcaggc tatgaggaag 2040

gccctgaccc tgaggtatgc cctgctgccc catctgtaca ccctgttcca ccaggcccat 2100

gtggctgggg agactgtggc caggcccctg tttctggagt ttcccaagga cagcagcacc 2160

tggactgtgg accatcagct gctgtggggg gaggctctgc tgattacccc tgtgctgcag 2220

gctggcaagg ctgaggtgac tgggtacttc cccctgggga cttggtatga cctgcagact 2280

gtgcctgtgg aagctctggg cagcctgccc ccaccccctg ctgcccctag ggagcctgcc 2340

atccactctg agggccagtg ggtgaccctg cctgcccctc tggacaccat caatgtgcac 2400

ctgaggctg gctatatcat ccccctgcag ggccctgggc tgaccaccac tgagagcagg 2460

cagcagccca tggccctggc tgtggccctg actaaggggg gggaggccag gggggagctg 2520

ttctgggatg atggggagag cctggaggtg ctggagagag gggcctacac ccaggtgatc 2580

tttctggcca ggaacaacac cattgtgaat gagctggtga gggtgacttc tgagggggct 2640

ggcctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgcccccca gcaggtgctg 2700

agcaatgggg tgcctgtgtc taacttcacc tacagccctg atactaaggt gctggatatc 2760

tgtgtgagcc tgctgatggg ggagcagttt ctggtgagct ggtgctgaag atctagagct 2820

gaattcctgc agccagggg atcagcctct actgtgcctt ctagttgcca gccatctgtt 2880

gtttgcccct cccccttgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 2940

taataaaatg aggaaattgc atcacattgt ctgagtaggt gtcattctat tctggggggt 3000

ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 3060

gcagtgggct ctatgg 3076

<210> 15

<211> 3076

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 15

atggcctttc tgtggctgct gtcctgctgg gccctgctgg ggaccacctt tggcctgctg 60

gtgcccagg agctgtctgg gagcagccca gtgctggagg agacccaccc tgcccaccag 120

caggggggcca gcaggcctgg ccctagggat gcccaggccc accctggcag gcccaggggct 180

gtgcctaccc agtgtgatgt gccacccaat tctaggtttg actgtgctcc tgacaaggcc 240

atcactcagg agcagtgtga agctaggggg tgctgctaca tcccagccaa gcagggcctg 300

caggggggccc agatgggcca gccctggtgc ttcttccccc ccagctaccc tagctacaag 360

ctggagaatc tgagcagctc tgagatgggc tacactgcta ccctgaccag gaccactcct 420

accttcttcc ccaaggacat cctgactctg aggctggatg tcatgatgga gactgaaaat 480

aggctgcact tcaccatcaa ggaccctgcc aataggaggt atgaggtgcc tctggagacc 540

ccccatgtgc atagcagggc tcccagcccc ctgtattctg tggagttctc tgaggagccc 600

tttggggtca ttgtgaggag acagctggat gggagggtgc tgctgaacac tactgtggct 660

cccctgttct ttgctgacca gttcctgcag ctgtctacca gcctgcccag ccagtacatc 720

actgggctgg ctgagcatct gagccccctg atgctgagca ccagctggac caggatcact 780

ctgtggaaca gggatctggc ccccactcct ggggccaacc tgtatgggag ccatcccttc 840

tacctggccc tggaggatgg gggctctgcc catggggtgt tcctgctgaa cagcaatgcc 900

atggatgtgg tgctgcagcc tagccctgcc ctgagctgga ggagcactgg gggcatcctg 960

gatgtctaca tcttcctggg gcctgagccc aagtctgtgg tgcagcagta tctggatgtg 1020

gtggggtatc ccttcatgcc cccctactgg ggcctgggct ttcacctgtg caggtggggc 1080

tacagcagca ctgccatcac caggcaggtg gtggagaaca tgaccagggc ccacttccct 1140

ctggatgtgc agtggaatga cctggactat atggattcta gggagagactt tacttttaac 1200

aaggatggct tcagggattt ccctgccatg gtgcaggagc tgcaccaggg gggcaggagg 1260

tacatgatga ttgtggaccc tgctattagc agctctggcc ctgctgggtc ttacaggcct 1320

tatgatgagg gcctgaggag gggggtgttc atcaccaatg agactggcca gcccctgatt 1380

ggcaaggtgt ggcctggcag cactgccttc cctgacttca ccaaccccac tgccctggcc 1440

tggtgggagg acatggtggc tgagttccat gaccaggtgc cctttgatgg gatgtggatt 1500

gacatgaatg agccctctaa cttcatcagg gggtctgagg atggctgccc caacaatgag 1560

ctggagaacc ccccctatgt gcctggggtg gtggggggca ctctgcaggc tgccactatc 1620

tgtgcttctt ctcaccagtt tctgagcacc cactataatc tgcacaacct gtatggcctg 1680

actgaggcca ttgccagcca tagggccctg gtgaaggcca ggggcaccag gccctttgtg 1740

atcagcaggt ctacctttgc tggccatggc aggtatgctg gccactggac tggggatgtg 1800

tggtcttctt ggggagcagct ggccagctct gtgcctgaga tcctgcagtt caacctgctg 1860

ggggtgcctc tggtgggggc tgatgtgtgt ggctttctgg gcaacacctc tgaggagctg 1920

tgtgtgaggt ggacccagct gggggccttt taccccttca tgaggaacca caatagcctg 1980

ctgagcctgc cccaggagcc ttactctttc tctgagcctg cccagcaggc catgaggaag 2040

gccctgactc tgaggtatgc cctgctgccc catctgtata ccctgtttca ccaggcccat 2100

gtggctgggg agactgtggc taggcctctg tttctggagt tccctaagga ctctagcacc 2160

tggactgtgg accaccagct gctgtggggg gaggccctgc tgatcacccc tgtgctgcag 2220

gctggcaagg ctgaggtgac tggctacttc cccctgggca cctggtatga cctgcagact 2280

gtgcctgtgg aggccctggg gagcctgcct cccccccctg ctgcccccag ggagcctgcc 2340

attcattctg agggccagtg ggtgaccctg cctgcccctc tggacaccat caatgtgcac 2400

ctgaggctg ggtacatcat ccccctgcag ggccctggcc tgaccaccac tgagagcagg 2460

cagcagccca tggccctggc tgtggctctg accaaggggg gggaggccag gggggagctg 2520

ttctgggatg atggggagtc tctggaggtg ctggagaggg gggcctacac ccaggtgatc 2580

tttctggcca ggaacaatac tattgtgaat gagctggtga gggtgacctc tgagggggct 2640

ggcctgcagc tgcagaaggt gactgtgctg ggggtggcca ctgcccccca gcaggtcctg 2700

agcaatgggg tgcctgtgag caacttcacc tactctcctg acaccaaggt gctggacatt 2760

tgtgtgtctc tgctgatggg ggagcagttc ctggtgagct ggtgctgaag atctagagct 2820

gaattcctgc agccagggg atcagcctct actgtgcctt ctagttgcca gccatctgtt 2880

gtttgcccct cccccttgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 2940

taataaaatg aggaaattgc atcacattgt ctgagtaggt gtcattctat tctggggggt 3000

ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 3060

gcagtgggct ctatgg 3076

<210> 16

<211> 4730

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 16

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggcctag taggctcaga ggcacacagg agtttctggg 180

ctcaccctgc ccccttccaa cccctcagtt cccatcctcc agcagctgtt tgtgtgctgc 240

ctctgaagtc cacactgaac aaacttcagc ctactcatgt ccctaaaatg ggcaaacatt 300

gcaagcagca aacagcaaac acacagccct ccctgcctgc tgaccttgga gctggggcag 360

aggtcagaga cctctctggg cccatgccac ctccaacatc cactcgaccc cttggaattt 420

cggtggagag gagcagaggt tgtcctggcg tggtttaggt agtgtgagag gggtacccgg 480

ggatcttgct accagtggaa cagccactaa ggattctgca gtgagagcag agggccagct 540

aagtggtact ctcccagaga ctgtctgact cacgccaccc cctccacctt ggacacagga 600

cgctgtggtt tctgagccag gtacaatgac tcctttcggt aagtgcagtg gaagctgtac 660

actgcccagg caaagcgtcc gggcagcgta ggcgggcgac tcagatccca gccagtggac 720

ttagcccctg tttgctcctc cgataactgg ggtgaccttg gttaatattc accagcagcc 780

tcccccgttg cccctctgga tccactgctt aaatacggac gaggacaggg ccctgtctcc 840

tcagcttcag gcaccaccac tgacctggga cagtgaatac cactttcaca atctgctagc 900

gtttaaacga tcctgagaac ttcagggtga gtctatggga cccttgatgt tttctttccc 960

cttcttttct atggttaagt tcatgtcata ggaaggggag aagtaacagg gtacacatat 1020

tgaccaaatc agggtaattt tgcatttgta attttaaaaa atgctttctt cttttaatat 1080

acttttttgt ttatcttatt tctaatactt tccctaatct ctttctttca gggcaataat 1140

gatacaatgt atcatgcctc tttgcaccat tctaaagaat aacagtgata atttctgggt 1200

taaggcaata gcaatatttc tgcatataaa tatttctgca tataaattgt aactgatgta 1260

agaggtttca tattgctaat agcagctaca atccagctac cattctgctt ttattttctg 1320

gttgggataa ggctggatta ttctgagtcc aagctaggcc cttttgctaa tcttgttcat 1380

acctcttatc ttcctcccac agctcctggg caacctgctg gtctctctgc tggcccatca 1440

ctttggcaaa gcacgcgtgc caccatggcc tttctgtggc tgctgtcctg ctgggccctg 1500

ctggggacca cctttggcct gctggtgccc agggagctgt ctgggagcag cccagtgctg 1560

gaggaaccc accctgccca ccagcagggg gccagcaggc ctggccctag ggatgcccag 1620

gcccaccctg gcaggcccag ggctgtgcct acccagtgtg atgtgccacc caattctagg 1680

tttgactgtg ctcctgacaa ggccatcact caggagcagt gtgaagctag ggggtgctgc 1740

tacatcccag ccaagcaggg cctgcagggg gccgatgg gccagccctg gtgcttcttc 1800

ccccccagct accctagcta caagctggag aatctgagca gctctgagat gggctacact 1860

gctaccctga ccaggaccac tcctaccttc ttccccaagg acatcctgac tctgaggctg 1920

gatgtcatga tggagactga aaataggctg cacttcacca tcaaggaccc tgccaatagg 1980

aggtatgagg tgcctctgga gaccccccat gtgcatagca gggctcccag ccccctgtat 2040

tctgtggagt tctctgagga gccctttggg gtcattgtga ggagacagct ggatgggagg 2100

gtgctgctga acactactgt ggctcccctg ttctttgctg accagttcct gcagctgtct 2160

accagcctgc ccagccagta catcactggg ctggctgagc atctgagccc cctgatgctg 2220

agcaccagct ggaccaggat cactctgtgg aacagggatc tggcccccac tcctggggcc 2280

aacctgtatg ggagccatcc cttctacctg gccctggagg atgggggctc tgcccatggg 2340

gtgttcctgc tgaacagcaa tgccatggat gtggtgctgc agcctagccc tgccctgagc 2400

tggagggagca ctggggggcat cctggatgtc tacatcttcc tggggcctga gcccaagtct 2460

gtggtgcagc agtatctgga tgtggtgggg tatcccttca tgccccccta ctggggcctg 2520

ggctttcacc tgtgcaggtg gggctacagc agcactgcca tcaccaggca ggtggtggag 2580

aacatgacca gggcccactt ccctctggat gtgcagtgga atgacctgga ctatatggat 2640

tctaggagag actttacttt taacaaggat ggcttcaggg atttccctgc catggtgcag 2700

gagctgcacc aggggggcag gaggtacatg atgattgtgg accctgctat tagcagctct 2760

ggccctgctg ggtcttacag gccttatgat gagggcctga ggaggggggt gttcatcacc 2820

aatgagactg gccagcccct gattggcaag gtgtggcctg gcagcactgc cttccctgac 2880

ttcaccaacc ccactgccct ggcctggtgg gaggacatgg tggctgagtt ccatgaccag 2940

gtgccctttg atgggatgtg gattgacatg aatgagccct ctaacttcat caggggggtct 3000

gaggatggct gccccaacaa tgagctggag aaccccccct atgtgcctgg ggtggtgggg 3060

ggcactctgc aggctgccac tatctgtgct tcttctcacc agtttctgag cacccactat 3120

aatctgcaca acctgtatgg cctgactgag gccattgcca gccatagggc cctggtgaag 3180

gccaggggca ccaggccctt tgtgatcagc aggtctacct ttgctggcca tggcaggtat 3240

gctggccact ggactgggga tgtgtggtct tcttgggagc agctggccag ctctgtgcct 3300

gagatcctgc agttcaacct gctggggggtg cctctggtgg gggctgatgt gtgtggcttt 3360

ctgggcaaca cctctgagga gctgtgtgtg aggtggaccc agctgggggc cttttacccc 3420

ttcatgagga accacaatag cctgctgagc ctgccccagg agccttactc tttctctgag 3480

cctgcccagc aggccatgag gaaggccctg actctgaggt atgccctgct gccccatctg 3540

tataccctgt ttcaccaggc ccatgtggct ggggagactg tggctaggcc tctgtttctg 3600

gagttcccta aggactctag cacctggact gtggaccacc agctgctgtg gggggaggcc 3660

ctgctgatca cccctgtgct gcaggctggc aaggctgagg tgactggcta cttccccctg 3720

ggcacctggt atgacctgca gactgtgcct gtggaggccc tggggagcct gcctcccccc 3780

cctgctgccc ccagggagcc tgccattcat tctgagggcc agtgggtgac cctgcctgcc 3840

cctctggaca ccatcaatgt gcacctgagg gctgggtaca tcatccccct gcagggccct 3900

ggcctgacca ccactgagag caggcagcag cccatggccc tggctgtggc tctgaccaag 3960

gggggggagg ccagggggga gctgttctgg gatgatgggg agtctctgga ggtgctggag 4020

aggggggcct acacccaggt gatctttctg gccaggaaca atactattgt gaatgagctg 4080

gtgaggtga cctctgaggg ggctggcctg cagctgcaga aggtgactgt gctggggggtg 4140

gccactgccc cccagcaggt cctgagcaat ggggtgcctg tgagcaactt cacctactct 4200

cctgacacca aggtgctgga catttgtgtg tctctgctga tgggggagca gttcctggtg 4260

agctggtgct gaagatctag agctgaattc ctgcagccag ggggatcagc ctctactgtg 4320

ccttctagtt gccagccatc tgttgtttgc ccctccccct tgccttcctt gaccctggaa 4380

ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcaca ttgtctgagt 4440

aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 4500

gacaatagca ggcatgctgg ggatgcagtg ggctctatgg cttctgaggc agaaagaacc 4560

agctggggct cgagatccac tagggccgca ggaaccccta gtgatggagt tggccactcc 4620

ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg 4680

ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg 4730

<210> 17

<211> 4701

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 17

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggcctag taggctcaga ggcacacagg agtttctggg 180

ctcaccctgc ccccttccaa cccctcagtt cccatcctcc agcagctgtt tgtgtgctgc 240

ctctgaagtc cacactgaac aaacttcagc ctactcatgt ccctaaaatg ggcaaacatt 300

gcaagcagca aacagcaaac acacagccct ccctgcctgc tgaccttgga gctggggcag 360

aggtcagaga cctctctggg cccatgccac ctccaacatc cactcgaccc cttggaattt 420

cggtggagag gagcagaggt tgtcctggcg tggtttaggt agtgtgagag gggtacccgg 480

ggatcttgct accagtggaa cagccactaa ggattctgca gtgagagcag agggccagct 540

aagtggtact ctcccagaga ctgtctgact cacgccaccc cctccacctt ggacacagga 600

cgctgtggtt tctgagccag gtacaatgac tcctttcggt aagtgcagtg gaagctgtac 660

actgcccagg caaagcgtcc gggcagcgta ggcgggcgac tcagatccca gccagtggac 720

ttagcccctg tttgctcctc cgataactgg ggtgaccttg gttaatattc accagcagcc 780

tcccccgttg cccctctgga tccactgctt aaatacggac gaggacaggg ccctgtctcc 840

tcagcttcag gcaccaccac tgacctggga cagtgaatag atcctgagaa cttcagggtg 900

agtctatggg acccttgatg ttttctttcc ccttcttttc tatggttaag ttcatgtcat 960

aggaagggga gaagtaacag ggtacacata ttgaccaaat cagggtaatt ttgcatttgt 1020

aattttaaaa aatgctttct tcttttaata tacttttttg tttatcttat ttctaatact 1080

ttccctaatc tctttctttc agggcaataa tgatacaatg tatcatgcct ctttgcacca 1140

ttctaaagaa taacagtgat aatttctggg ttaaggcaat agcaatattt ctgcatataa 1200

atatttctgc atataaattg taactgatgt aagaggtttc atattgctaa tagcagctac 1260

aatccagcta ccattctgct tttattttct ggttgggata aggctggatt attctgagtc 1320

caagctaggc ccttttgcta atcttgttca tacctcttat cttcctccca cagctcctgg 1380

gcaacctgct ggtctctctg ctggcccatc actttggcaa agcacgcgtg ccaccatggc 1440

ctttctgtgg ctgctgtcct gctgggccct gctggggacc acctttggcc tgctggtgcc 1500

cagggagctg tctgggagca gcccagtgct ggaggagacc caccctgccc accagcaggg 1560

ggccagcagg cctggcccta gggatgccca ggcccaccct ggcaggccca gggctgtgcc 1620

tacccagtgt gatgtgccac ccaattctag gtttgactgt gctcctgaca aggccatcac 1680

tcaggagcag tgtgaagcta gggggtgctg ctacatccca gccaagcagg gcctgcaggg 1740

ggccccagatg ggccagccct ggtgcttctt cccccccagc taccctagct acaagctgga 1800

gaatctgagc agctctgaga tgggctacac tgctaccctg accaggacca ctcctacctt 1860

cttccccaag gacatcctga ctctgaggct ggatgtcatg atggagactg aaaataggct 1920

gcacttcacc atcaaggacc ctgccaatag gaggtatgag gtgcctctgg agacccccca 1980

tgtgcatagc agggctccca gccccctgta ttctgtggag ttctctgagg agccctttgg 2040

ggtcattgtg aggagacagc tggatgggag ggtgctgctg aacactactg tggctcccct 2100

gttctttgct gaccagttcc tgcagctgtc taccagcctg cccagccagt acatcactgg 2160

gctggctgag catctgagcc ccctgatgct gagcaccagc tggaccagga tcactctgtg 2220

gaacagggat ctggccccca ctcctggggc caacctgtat gggagccatc ccttctacct 2280

ggccctggag gatggggggct ctgcccatgg ggtgttcctg ctgaacagca atgccatgga 2340

tgtggtgctg cagcctagcc ctgccctgag ctggagggagc actgggggca tcctggatgt 2400

ctacatcttc ctggggcctg agcccaagtc tgtggtgcag cagtatctgg atgtggtggg 2460

gtatcccttc atgcccccct actggggcct gggctttcac ctgtgcaggt ggggctacag 2520

cagcactgcc atcaccaggc aggtggtgga gaacatgacc agggcccact tccctctgga 2580

tgtgcagtgg aatgacctgg actatatgga ttctaggaga gactttactt ttaacaagga 2640

tggcttcagg gatttccctg ccatggtgca ggagctgcac caggggggca ggaggtacat 2700

gatgattgtg gaccctgcta ttagcagctc tggccctgct gggtcttaca ggccttatga 2760

tgaggggcctg aggagggggg tgttcatcac caatgagact ggccagcccc tgattggcaa 2820

ggtgtggcct ggcagcactg ccttccctga cttcaccaac cccactgccc tggcctggtg 2880

ggaggacatg gtggctgagt tccatgacca ggtgcccttt gatgggatgt ggattgacat 2940

gaatgagccc tctaacttca tcagggggtc tgaggatggc tgccccaaca atgagctgga 3000

gaaccccccc tatgtgcctg gggtggtggg gggcactctg caggctgcca ctatctgtgc 3060

ttcttctcac cagtttctga gcacccacta taatctgcac aacctgtatg gcctgactga 3120

ggccattgcc agccataggg ccctggtgaa ggccaggggc accaggccct ttgtgatcag 3180

caggtctacc tttgctggcc atggcaggta tgctggccac tggactgggg atgtgtggtc 3240

ttcttgggag cagctggcca gctctgtgcc tgagatcctg cagttcaacc tgctgggggt 3300

gcctctggtg ggggctgatg tgtgtggctt tctgggcaac acctctgagg agctgtgtgt 3360

gaggtggacc cagctggggg ccttttaccc cttcatgagg aaccacaata gcctgctgag 3420

cctgccccag gagccttact ctttctctga gcctgcccag caggccatga ggaaggccct 3480

gactctgagg tatgccctgc tgccccatct gtataccctg tttcaccagg cccatgtggc 3540

tggggagact gtggctaggc ctctgtttct ggagttccct aaggactcta gcacctggac 3600

tgtggaccac cagctgctgt ggggggaggc cctgctgatc acccctgtgc tgcaggctgg 3660

caaggctgag gtgactggct acttccccct gggcacctgg tatgacctgc agactgtgcc 3720

tgtggaggcc ctggggagcc tgcctccccc ccctgctgcc cccagggagc ctgccattca 3780

ttctgaggc cagtgggtga ccctgcctgc ccctctggac accatcaatg tgcacctgag 3840

ggctgggtac atcatccccc tgcagggccc tggcctgacc accactgaga gcaggcagca 3900

gcccatggcc ctggctgtgg ctctgaccaa ggggggggag gccagggggg agctgttctg 3960

ggatgatggg gagtctctgg aggtgctgga gaggggggcc tacacccagg tgatctttct 4020

ggccaggaac aatactattg tgaatgagct ggtgaggtg acctctgagg gggctggcct 4080

gcagctgcag aaggtgactg tgctgggggt ggccactgcc ccccagcagg tcctgagcaa 4140

tggggtgcct gtgagcaact tcacctactc tcctgacacc aaggtgctgg acatttgtgt 4200

gtctctgctg atgggggagc agttcctggt gagctggtgc tgaagatcta gagctgaatt 4260

cctgcagcca gggggatcag cctctactgt gccttctagt tgccagccat ctgttgtttg 4320

cccctccccc ttgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 4380

aaatgaggaa attgcatcac attgtctgag taggtgtcat tctattctgg ggggtggggt 4440

ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcagt 4500

gggctctatg gcttctgagg cagaaagaac cagctggggc tcgagatcca ctagggccgc 4560

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 4620

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 4680

gagcgcgcag ctgcctgcag g 4701

<210> 18

<211> 4701

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 18

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggcctag taggctcaga ggcacacagg agtttctggg 180

ctcaccctgc ccccttccaa cccctcagtt cccatcctcc agcagctgtt tgtgtgctgc 240

ctctgaagtc cacactgaac aaacttcagc ctactcatgt ccctaaaatg ggcaaacatt 300

gcaagcagca aacagcaaac acacagccct ccctgcctgc tgaccttgga gctggggcag 360

aggtcagaga cctctctggg cccatgccac ctccaacatc cactcgaccc cttggaattt 420

cggtggagag gagcagaggt tgtcctggcg tggtttaggt agtgtgagag gggtacccgg 480

ggatcttgct accagtggaa cagccactaa ggattctgca gtgagagcag agggccagct 540

aagtggtact ctcccagaga ctgtctgact cacgccaccc cctccacctt ggacacagga 600

cgctgtggtt tctgagccag gtacaatgac tcctttcggt aagtgcagtg gaagctgtac 660

actgcccagg caaagcgtcc gggcagcgta ggcgggcgac tcagatccca gccagtggac 720

ttagcccctg tttgctcctc cgataactgg ggtgaccttg gttaatattc accagcagcc 780

tcccccgttg cccctctgga tccactgctt aaatacggac gaggacaggg ccctgtctcc 840

tcagcttcag gcaccaccac tgacctggga cagtgaatag atcctgagaa cttcagggtg 900

agtctatggg acccttgatg ttttctttcc ccttcttttc tatggttaag ttcatgtcat 960

aggaagggga gaagtaacag ggtacacata ttgaccaaat cagggtaatt ttgcatttgt 1020

aattttaaaa aatgctttct tcttttaata tacttttttg tttatcttat ttctaatact 1080

ttccctaatc tctttctttc agggcaataa tgatacaatg tatcatgcct ctttgcacca 1140

ttctaaagaa taacagtgat aatttctggg ttaaggcaat agcaatattt ctgcatataa 1200

atatttctgc atataaattg taactgatgt aagaggtttc atattgctaa tagcagctac 1260

aatccagcta ccattctgct tttattttct ggttgggata aggctggatt attctgagtc 1320

caagctaggc ccttttgcta atcttgttca tacctcttat cttcctccca cagctcctgg 1380

gcaacctgct ggtctctctg ctggcccatc actttggcaa agcacgcgtg ccaccatggc 1440

tttcctgtgg ctgctgtctt gttgggctct gctgggcacc acctttggcc tgctggtgcc 1500

cagggagctg tctggcagca gccctgtgct ggagaggacc caccctgctc atcagcaggg 1560

ggctagcagg cctggcccca gggatgccca ggctcaccct gggaccca gggctgtgcc 1620

cactcagtgt gatgtgcccc ccaacagcag gtttgactgt gctcctgaca aggctatcac 1680

ccaggagcag tgtgaggcca gggggtgctg ctacattcct gctaagcagg gcctgcaggg 1740

ggcccagatg ggccagccct ggtgcttctt ccccccctct tatcccagct ataagctgga 1800

gaacctgagc agctctgaga tgggctacac tgccaccctg accaggacca ctcccacctt 1860

ctttcccaag gatattctga ctctgaggct ggatgtgatg atggagactg agaacaggct 1920

gcacttcact atcaaggacc ctgccaatag gaggtatgag gtgcccctgg agactcctca 1980

tgtgcatagc agggcccctt ctcctctgta ttctgtggag ttctctgagg agccctttgg 2040

ggtgattgtg aggaggcagc tggatggcag ggtgctgctg aacaccactg tggcccccct 2100

gttctttgct gaccagttcc tgcagctgag cacttctctg cccagccagt acattactgg 2160

gctggctgag catctgagcc ccctgatgct gagcacctct tggaccagga tcaccctgtg 2220

gaacagggac ctggccccca ctcctggggc taacctgtat ggctctcacc ccttttacct 2280

ggccctggag gatggggggct ctgcccatgg ggtgtttctg ctgaacagca atgctatgga 2340

tgtggtgctg cagccctctc cagccctgtc ttggagggagc actggggggca ttctggatgt 2400

gtacattttc ctggggcctg aacccaagtc tgtggtgcag cagtacctgg atgtggtggg 2460

ctaccccttc atgcccccct attggggggct ggggtttcac ctgtgcaggt ggggctacag 2520

cagcactgcc atcaccaggc aggtggtgga gaacatgacc agggcccatt tccccctgga 2580

tgtgcagtgg aatgacctgg actacatgga tagcaggagg gatttcacct tcaacaagga 2640

tggcttcagg gactttcctg ccatggtgca ggagctgcac caggggggca ggaggtatat 2700

gatgattgtg gaccctgcta tcagcagctc tggccctgct ggctcttaca ggccctatga 2760

tgaggggcctg aggagggggg tgtttatcac taatgaaact ggccagcctc tgattggcaa 2820

ggtctggcct ggctctactg ccttccctga ttttactaac cccactgccc tggcctggtg 2880

ggaggacatg gtggctgagt tccatgatca ggtgcctttt gatggcatgt ggattgatat 2940

gaatgaacca agcaacttca tcagaggctc tgaggatggc tgccccaaca atgagctgga 3000

gaaccccccc tatgtgcctg gggtggtggg gggcactctg caggctgcca ccatttgtgc 3060

tagcagccac cagttcctga gcacccacta caatctgcac aacctgtatg gcctgactga 3120

agccattgcc agccataggg ccctggtgaa ggccaggggc actaggcctt ttgtgatcag 3180

caggagcact tttgctggcc atggcaggta tgctggccac tggactgggg atgtgtggag 3240

cagctgggag cagctggcca gctctgtgcc tgagattctg cagtttaacc tgctgggggt 3300

gcccctggtg ggggctgatg tgtgtggctt cctgggcaac acctctgagg agctgtgtgt 3360

gaggtggacc cagctggggg ccttttatcc cttcatgagg aaccacaaca gcctgctgag 3420

cctgcctcag gagccctact ctttctctga gcctgcccag caggccatga ggaaggccct 3480

gaccctgagg tatgccctgc tgccccacct gtataccctg ttccaccagg cccatgtggc 3540

tggggagact gtggccaggc ccctgttcct ggagttcccc aaggacagca gcacctggac 3600

tgtggatcat cagctgctgt ggggggaggc cctgctgatc acccctgtgc tgcaggctgg 3660

caaggctgag gtcactggct acttccctct gggcacctgg tatgacctgc agactgtgcc 3720

tgtggaggct ctgggcagcc tgcccccccc ccctgctgct cccagggagc ctgccatcca 3780

ctctgagggc cagtgggtga ccctgcctgc tcccctggac accatcaatg tgcacctgag 3840

ggctggctac attatccccc tgcagggccc agggctgact accactgaga gcagacagca 3900

gcccatggct ctggctgtgg ccctgaccaa ggggggggaa gctagggggg agctgttctg 3960

ggatgatggg gagagcctgg aggtgctgga gaggggggcc tatacccagg tgatcttcct 4020

ggctaggaac aacaccattg tcaatgagct ggtgaggtg acttctgagg gggctgggct 4080

gcagctgcag aaggtgactg tgctgggggt ggccactgct ccccagcagg tgctgagcaa 4140

tggggtgcct gtgagcaact tcacctacag ccctgacacc aaggtgctgg acatctgtgt 4200

gtctctgctg atgggggagc agttcctggt gagctggtgc tgaagatcta gagctgaatt 4260

cctgcagcca gggggatcag cctctactgt gccttctagt tgccagccat ctgttgtttg 4320

cccctccccc ttgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 4380

aaatgaggaa attgcatcac attgtctgag taggtgtcat tctattctgg ggggtggggt 4440

ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcagt 4500

gggctctatg gcttctgagg cagaaagaac cagctggggc tcgagatcca ctagggccgc 4560

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 4620

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 4680

gagcgcgcag ctgcctgcag g 4701

<210> 19

<211> 4701

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 19

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggcctag taggctcaga ggcacacagg agtttctggg 180

ctcaccctgc ccccttccaa cccctcagtt cccatcctcc agcagctgtt tgtgtgctgc 240

ctctgaagtc cacactgaac aaacttcagc ctactcatgt ccctaaaatg ggcaaacatt 300

gcaagcagca aacagcaaac acacagccct ccctgcctgc tgaccttgga gctggggcag 360

aggtcagaga cctctctggg cccatgccac ctccaacatc cactcgaccc cttggaattt 420

cggtggagag gagcagaggt tgtcctggcg tggtttaggt agtgtgagag gggtacccgg 480

ggatcttgct accagtggaa cagccactaa ggattctgca gtgagagcag agggccagct 540

aagtggtact ctcccagaga ctgtctgact cacgccaccc cctccacctt ggacacagga 600

cgctgtggtt tctgagccag gtacaatgac tcctttcggt aagtgcagtg gaagctgtac 660

actgcccagg caaagcgtcc gggcagcgta ggcgggcgac tcagatccca gccagtggac 720

ttagcccctg tttgctcctc cgataactgg ggtgaccttg gttaatattc accagcagcc 780

tcccccgttg cccctctgga tccactgctt aaatacggac gaggacaggg ccctgtctcc 840

tcagcttcag gcaccaccac tgacctggga cagtgaatag atcctgagaa cttcagggtg 900

agtctatggg acccttgatg ttttctttcc ccttcttttc tatggttaag ttcatgtcat 960

aggaagggga gaagtaacag ggtacacata ttgaccaaat cagggtaatt ttgcatttgt 1020

aattttaaaa aatgctttct tcttttaata tacttttttg tttatcttat ttctaatact 1080

ttccctaatc tctttctttc agggcaataa tgatacaatg tatcatgcct ctttgcacca 1140

ttctaaagaa taacagtgat aatttctggg ttaaggcaat agcaatattt ctgcatataa 1200

atatttctgc atataaattg taactgatgt aagaggtttc atattgctaa tagcagctac 1260

aatccagcta ccattctgct tttattttct ggttgggata aggctggatt attctgagtc 1320

caagctaggc ccttttgcta atcttgttca tacctcttat cttcctccca cagctcctgg 1380

gcaacctgct ggtctctctg ctggcccatc actttggcaa agcacgcgtg ccaccatggc 1440

cttcctgtgg ctgctgtctt gctgggctct gctggggacc acctttggcc tgctggtccc 1500

cagggagctg tctggctctt ctcctgtcct ggagaggacc caccctgccc accagcaggg 1560

ggctagcagg cctggcccca gggatgccca ggcccaccct ggcaggccca gggctgtgcc 1620

cacccagtgt gatgtgcctc ccaacagcag gtttgactgt gcccctgaca aggccatcac 1680

ccaggagcag tgtgaggcca ggggctgctg ctatatccct gccaagcagg gcctgcaggg 1740

ggctcagatg ggccagccct ggtgcttctt tcccccctct tatcctagct ataagctgga 1800

gaacctgagc agctctgaga tggggtacac tgccaccctg accaggacca cccccacttt 1860

cttccctaag gacatcctga ccctgaggct ggatgtgatg atggagactg agaataggct 1920

gcactttact atcaaggacc ctgccaacag gaggtatgag gtgcctctgg agacccccca 1980

tgtgcattct agggccccca gccccctgta ctctgtggag ttctctgagg agccctttgg 2040

ggtgattgtg aggagacagc tggatggcag ggtcctgctg aacaccactg tggctcccct 2100

gttttttgct gaccagttcc tgcagctgag caccagcctg cccagccagt acatcactgg 2160

gctggctgag cacctgagcc ccctgatgct gagcaccagc tggaccagga tcaccctgtg 2220

gaacagggat ctggctccta cccctggggc caacctgtat ggctctcacc ccttttacct 2280

ggccctggag gatggggggct ctgcccatgg ggtgttcctg ctgaacagca atgctatgga 2340

tgtggtgctg cagcccagcc ctgccctgag ctggaggtct actgggggca tcctggatgt 2400

gtacatcttt ctggggcctg agcccaagtc tgtggtgcag cagtacctgg atgtggtggg 2460

ctatcctttt atgcccccct attggggcct gggcttccac ctgtgcaggt ggggctacag 2520

cagcactgcc atcaccagac aggtggtgga gaacatgacc agggcccact tccccctgga 2580

tgtgcagtgg aatgacctgg actacatgga cagcaggagg gacttcacct ttaacaagga 2640

tggctttagg gacttccctg ccatggtgca ggagctgcat caggggggca ggaggtacat 2700

gatgattgtg gacccagcca tcagcagctc tgggcctgct gggtcttaca ggccctatga 2760

tgaggggcctg aggagggggg tgttcatcac caatgagact ggccagcccc tgattggcaa 2820

ggtgtggcct gggagcactg ccttccctga ttttaccaac cccactgccc tggcctggtg 2880

ggaggatatg gtggctgagt ttcatgacca ggtgcccttt gatggcatgt ggattgacat 2940

gaatgagccc agcaatttca tcaggggctc tgaggatggc tgccccaaca atgagctgga 3000

gaatcctccc tatgtgcctg gggtggtggg gggcaccctg caggctgcca ccatctgtgc 3060

ctctagccac cagttcctga gcacccacta taacctgcat aacctgtatg gcctgactga 3120

ggccattgcc agccatagag ccctggtgaa ggccagaggg accaggccct ttgtgatctc 3180

taggagcacc tttgctggcc atggcaggta tgctggccac tggactgggg atgtgtggag 3240

ctcttgggag cagctggcca gctctgtgcc agagatcctg cagttcaacc tgctgggggt 3300

gcctctggtg ggggctgatg tgtgtggctt cctgggcaat acctctgaag agctgtgtgt 3360

gaggtggact cagctggggg ccttctatcc cttcatgagg aaccacaaca gcctgctgtc 3420

tctgccccag gagccctaca gcttctctga gcctgctcag caggctatga ggaaggccct 3480

gaccctgagg tatgccctgc tgccccatct gtacaccctg ttccaccagg cccatgtggc 3540

tggggagact gtggccaggc ccctgtttct ggagtttccc aaggacagca gcacctggac 3600

tgtggaccat cagctgctgt ggggggaggc tctgctgatt acccctgtgc tgcaggctgg 3660

caaggctgag gtgactgggt acttccccct ggggacttgg tatgacctgc agactgtgcc 3720

tgtggaagct ctgggcagcc tgcccccacc ccctgctgcc cctagggagc ctgccatcca 3780

ctctgaggc cagtgggtga ccctgcctgc ccctctggac accatcaatg tgcacctgag 3840

ggctggctat atcatccccc tgcagggccc tgggctgacc accactgaga gcaggcagca 3900

gcccatggcc ctggctgtgg ccctgactaa ggggggggag gccagggggg agctgttctg 3960

ggatgatggg gagagcctgg aggtgctgga gagaggggcc tacacccagg tgatctttct 4020

ggccaggaac aacaccattg tgaatgagct ggtgaggtg acttctgagg gggctggcct 4080

gcagctgcag aaggtgactg tgctgggggt ggccactgcc ccccagcagg tgctgagcaa 4140

tggggtgcct gtgtctaact tcacctacag ccctgatact aaggtgctgg atatctgtgt 4200

gagcctgctg atgggggagc agtttctggt gagctggtgc tgaagatcta gagctgaatt 4260

cctgcagcca gggggatcag cctctactgt gccttctagt tgccagccat ctgttgtttg 4320

cccctccccc ttgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 4380

aaatgaggaa attgcatcac attgtctgag taggtgtcat tctattctgg ggggtggggt 4440

ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcagt 4500

gggctctatg gcttctgagg cagaaagaac cagctggggc tcgagatcca ctagggccgc 4560

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 4620

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 4680

gagcgcgcag ctgcctgcag g 4701

<210> 20

<211> 4611

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 20

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggccgca ggctcagagg cacacaggag tttctgggct 180

caccctgccc ccttccaacc cctcagttcc catcctccag cagctgtttg tgtgctgcct 240

ctgaagtcca cactgaacaa acttcagcct actcatgtcc ctaaaatggg caaacattgc 300

aagcagcaaa cagcaaacac acagccctcc ctgcctgctg accttggagc tggggcagag 360

gtcagagacc tctctgggcc catgccacct ccaacatcca ctcgacccct tggaatttcg 420

gtggagagga gcagaggttg tcctggcgtg gtttaggtag tgtgagaggg gtacccgggg 480

atcttgctac cagtggaaca gccactaagg attctgcagt gagagcagag ggccagctaa 540

gtggtactct cccagagact gtctgactca cgccaccccc tccaccttgg acacaggacg 600

ctgtggtttc tgagccaggt acaatgactc ctttcggtaa gtgcagtgga agctgtacac 660

tgcccaggca aagcgtccgg gcagcgtagg cgggcgactc agatcccagc cagtggactt 720

agcccctgtt tgctcctccg ataactgggg tgaccttggt taatattcac cagcagcctc 780

ccccgttgcc cctctggatc cactgcttaa atacggacga ggacagggcc ctgtctcctc 840

agcttcaggc accaccactg acctgggaca gtgaatagat cctgagaact tcagggtgag 900

tctatgggac ccttgatgtt ttctttcccc ttcttttcta tggttaagtt catgtcatag 960

gaaggggaga agtaacaggg tacacatatt gaccaaatca gggtaatttt gcatttgtaa 1020

ttttaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt ctaatacttt 1080

ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct ttgcaccatt 1140

ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct gcatataaat 1200

atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata gcagctacaa 1260

tccagctacc attctgcttt tattttctgg ttgggataag gctggattat tctgagtcca 1320

agctaggccc ttttgctaat cttgttcata cctcttatct tcctcccaca gctcctgggc 1380

aacctgctgg tctctctgct ggcccatcac tttggcaaag cacgcgtgcc accatggctt 1440

tcctgtggct gctgagctgc tgggctctgc tgggcaccac ctttgggctg ctggtgccta 1500

gggagctgtc tgggtctagc cctgtgctgg aggagactca ccctgcccat cagcagggg 1560

ctagcaggcc tggccccagg gatgctcagg cccaccctgg caggcccagg gctgtgccca 1620

cccagtgtga tgtgcccccc aacagcaggt ttgactgtgc ccctgacaag gccattaccc 1680

aggagcagtg tgaggccagg ggctgctgct acattccagc taagcagggc ctgcaggggg 1740

cccagatggg ccagccctgg tgcttcttcc cccccagcta tcctagctat aaactggaga 1800

acctgagcag ctctgagatg ggctatactg ccaccctgac taggactact cccacctttt 1860

ttcctaagga tatcctgacc ctgaggctgg atgtgatgat ggagactgag aacaggctgc 1920

acttcactat taaggaccct gccaatagga ggtatgaagt gcctctggag actcctcatg 1980

tgcactctag ggcccccagc cccctgtatt ctgtggagtt ctctgaggag ccctttgggg 2040

tgattgtgag gaggcagctg gatggcaggg tgctgctgaa caccactgtg gcccccctgt 2100

tctttgctga ccagttcctg cagctgagca ccagcctgcc cagccagtac atcactgggc 2160

tggctgagca tctgagccct ctgatgctga gcacctcttg gaccaggatc accctgtgga 2220

atagggatct ggcccccacc cctggggcta atctgtatgg ctctcatccc ttttacctgg 2280

ccctggagga tggggggctct gcccatgggg tgtttctgct gaacagcaat gccatggatg 2340

tggtgctgca gccctctcct gccctgagct ggaggagcac tgggggcatc ctggatgtgt 2400

acatcttcct gggccctgag cccaagtctg tggtccagca gtatctggat gtggtgggct 2460

acccctttat gcccccctat tggggcctgg gcttccacct gtgcaggtgg gggtattctt 2520

ctactgctat caccaggcag gtggtggaga acatgaccag ggctcacttc cccctggatg 2580

tgcagtggaa tgacctggac tatatggact ctaggaggga tttcaccttc aacaaggatg 2640

gcttcaggga cttccctgct atggtccagg agctgcatca ggggggcagg aggtacatga 2700

tgattgtgga ccctgccatc agcagctctg gccctgctgg cagctatagg ccctatgatg 2760

agggcctgag gaggggggtg tttatcacta atgaaactgg gcagcccctg attggcaagg 2820

tgtggcctgg ctctactgcc ttccctgact tcaccaaccc cactgctctg gcctggtggg 2880

aggacatggt ggctgagttc catgaccagg tgccttttga tggcatgtgg attgacatga 2940

atgagcccag caacttcatc aggggctctg aggatgggtg ccccaataat gagctggaga 3000

acccccccta tgtgcctggg gtggtggggg gcaccctgca ggctgccact atttgtgcca 3060

gctctcacca gttcctgagc accactaca acctgcacaa tctgtatggc ctgactgagg 3120

ccattgccag ccacaggggcc ctggtgaagg ccaggggcac taggcccttt gtgatctcta 3180

gaagcacctt tgctggccat gggaggtatg ctggccactg gactggggat gtgtggagct 3240

cttgggagca gctggccagc tctgtgcctg agatcctgca gttcaacctg ctggggggtgc 3300

ccctggtggg ggctgatgtg tgtggcttcc tgggcaacac ctctgaagag ctgtgtgtga 3360

ggtggaccca gctggggggcc ttctaccctt tcatgaggaa ccacaacagc ctgctgagcc 3420

tgcctcagga gccttactct ttctctgagc ctgcccagca ggccatgagg aaggccctga 3480

ccctgaggta tgctctgctg ccccacctgt acaccctgtt ccaccaggcc catgtggctg 3540

gggagactgt ggccaggccc ctgttcctgg agtttcctaa ggatagcagc acctggactg 3600

tggaccacca gctgctgtgg ggggaggccc tgctgattac ccctgtgctg caggctggca 3660

aggctgaggt gactggctac ttccccctgg gcacttggta tgacctgcag actgtgcctg 3720

tggaagccct gggcagcctg cctccccccc ctgctgcccc cagggagcct gccatccact 3780

ctgaggcca gtgggtgacc ctgcctgccc ccctggacac cattaatgtg catctgaggg 3840

ctgggtatat tatccccctg caggggcctg ggctgactac cactgagagc aggcagcagc 3900

ctatggccct ggctgtggct ctgactaagg ggggggaggc caggggggag ctgttctggg 3960

atgatgggga gagcctggag gtgctggaga ggggggccta cacccaggtg attttcctgg 4020

caggaacaa caccattgtg aatgagctgg tgaggtgac ctctgagggg gctggcctgc 4080

agctgcagaa agtgactgtg ctggggggtgg ccactgcccc ccagcaggtg ctgagcaatg 4140

gggtgcctgt gagcaacttc acctacagcc ctgacaccaa ggtgctggat atttgtgtga 4200

gcctgctgat gggggagcag ttcctggtga gctggtgctg actcgagaga tctaccggtg 4260

aattcaccgc gggtttaaac tgtgccttct agttgccagc catctgttgt ttgcccctcc 4320

cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 4380

gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtgggggct 4440

agctctagac tcgagatcca ctagggccgc aggaacccct agtgatggag ttggccactc 4500

cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 4560

gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag g 4611

<210> 21

<211> 4611

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 21

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggccgca ggctcagagg cacacaggag tttctgggct 180

caccctgccc ccttccaacc cctcagttcc catcctccag cagctgtttg tgtgctgcct 240

ctgaagtcca cactgaacaa acttcagcct actcatgtcc ctaaaatggg caaacattgc 300

aagcagcaaa cagcaaacac acagccctcc ctgcctgctg accttggagc tggggcagag 360

gtcagagacc tctctgggcc catgccacct ccaacatcca ctcgacccct tggaatttcg 420

gtggagagga gcagaggttg tcctggcgtg gtttaggtag tgtgagaggg gtacccgggg 480

atcttgctac cagtggaaca gccactaagg attctgcagt gagagcagag ggccagctaa 540

gtggtactct cccagagact gtctgactca cgccaccccc tccaccttgg acacaggacg 600

ctgtggtttc tgagccaggt acaatgactc ctttcggtaa gtgcagtgga agctgtacac 660

tgcccaggca aagcgtccgg gcagcgtagg cgggcgactc agatcccagc cagtggactt 720

agcccctgtt tgctcctccg ataactgggg tgaccttggt taatattcac cagcagcctc 780

ccccgttgcc cctctggatc cactgcttaa atacggacga ggacagggcc ctgtctcctc 840

agcttcaggc accaccactg acctgggaca gtgaatagat cctgagaact tcagggtgag 900

tctatgggac ccttgatgtt ttctttcccc ttcttttcta tggttaagtt catgtcatag 960

gaaggggaga agtaacaggg tacacatatt gaccaaatca gggtaatttt gcatttgtaa 1020

ttttaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt ctaatacttt 1080

ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct ttgcaccatt 1140

ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct gcatataaat 1200

atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata gcagctacaa 1260

tccagctacc attctgcttt tattttctgg ttgggataag gctggattat tctgagtcca 1320

agctaggccc ttttgctaat cttgttcata cctcttatct tcctcccaca gctcctgggc 1380

aacctgctgg tctctctgct ggcccatcac tttggcaaag cacgcgtgcc accatggctt 1440

tcctgtggct gctgtcttgc tgggccctgc tggggactac ctttggcctg ctggtgccca 1500

gggaactgtc tggctctagc ccagtgctgg aggagaccca ccctgcccac cagcagggg 1560

cttctaggcc tggccccagg gatgcccagg cccaccctgg caggccaagg gctgtgccca 1620

cccagtgtga tgtgcccccc aactctagat ttgattgtgc ccctgataag gccatcaccc 1680

aggagcagtg tgaggctagg ggctgctgct acatccctgc taagcagggc ctgcaggggg 1740

ctcagatggg ccagccctgg tgcttcttcc cccccagcta tccctcttac aagctggaga 1800

atctgagcag ctctgagatg ggctacactg ccaccctgac caggactact cccaccttct 1860

tccccaagga catcctgacc ctgaggctgg atgtgatgat ggagactgag aacaggctgc 1920

atttcaccat caaggatcct gccaacagga ggtatgaggt gcctctggag accccccatg 1980

tgcacagcag ggctccttct cccctgtact ctgtggagtt ctctgaggaa ccctttgggg 2040

tgattgtgag gaggcagctg gatggcaggg tcctgctgaa caccactgtg gcccccctgt 2100

tctttgctga tcagttcctg cagctgtcca cttctctgcc tagccagtac atcactgggc 2160

tggctgagca cctgagccct ctgatgctga gcacctcttg gactaggatc accctgtgga 2220

acagggacct ggcccccacc cctggggcca acctgtatgg cagccacccc ttctatctgg 2280

ccctggagga tggggggctct gcccatgggg tgttcctgct gaatagcaat gctatggatg 2340

tggtgctgca gcccagccct gccctgtctt ggagaggcac tgggggcatc ctggatgtgt 2400

acattttcct ggggcctgag cccaagtctg tggtgcagca gtacctggat gtggtgggct 2460

accccttcat gcctccctac tggggcctgg gcttccacct gtgcaggtgg ggctacagct 2520

ctactgccat caccaggcag gtggtggaga atatgaccag ggcccacttc cccctggatg 2580

tgcagtggaa tgacctggac tacatggact ctaggaggga cttcaccttc aataaggatg 2640

gcttcagaga cttccctgcc atggtgcagg agctgcatca ggggggcagg aggtacatga 2700

tgattgtgga ccctgccatc agctcttctg gccctgctgg ctcttacagg ccctatgatg 2760

agggcctgag gaggggggtg ttcatcacca atgagactgg gcagcccctg attgggaagg 2820

tgtggcctgg ctctactgcc ttccctgact tcaccaatcc tactgccctg gcctggtggg 2880

aggacatggt ggctgagttc catgaccagg tgccctttga tggcatgtgg attgacatga 2940

atgagccctc taatttcatc aggggctctg aggatggctg ccccaacaat gagctggaga 3000

acccccccta tgtgcctggg gtggtggggg gcaccctgca ggctgccacc atctgtgcta 3060

gctctcacca gttcctgagc acccactaca atctgcataa cctgtatggc ctgactgagg 3120

ccattgccag ccacaggggcc ctggtgaagg ctaggggcac caggcccttt gtgatttcta 3180

ggagcacttt tgctggccat ggcaggtatg ctgggcactg gactggggat gtgtggtcta 3240

gctgggagca gctggcttct tctgtgcctg agatcctgca gttcaacctg ctggggggtgc 3300

ctctggtggg ggctgatgtg tgtgggttcc tgggcaacac ttctgaggag ctgtgtgtga 3360

ggtggaccca gctggggggcc ttctaccctt tcatgaggaa ccacaacagc ctgctgagcc 3420

tgccccagga gccctacagc ttctctgagc ctgcccagca ggccatgagg aaggccctga 3480

ccctgaggta tgccctgctg ccccacctgt acaccctgtt ccaccaggcc catgtggctg 3540

gggagactgt ggctaggcct ctgttcctgg agttccccaa ggactctagc acctggactg 3600

tggaccacca gctgctgtgg ggggaggccc tgctgatcac tcctgtgctg caggctggga 3660

aggctgaggt gactggctat ttccccctgg gcacctggta tgacctgcag actgtgcctg 3720

tggaggccct ggggagcctg cccccccccc ctgctgcccc cagggagcct gccatccact 3780

ctgaggcca gtgggtgacc ctgcctgccc ctctggatac catcaatgtg cacctgaggg 3840

ctggctacat cattcccctg cagggccctg gcctgaccac tactgagtct aggcagcagc 3900

ccatggccct ggctgtggcc ctgaccaagg ggggggaggc taggggggag ctgttttggg 3960

atgatgggga gagcctggag gtgctggaga ggggggccta cactcaggtg atcttcctgg 4020

caggaacaa taccattgtg aatgagctgg tgaggtgac ctctgagggg gctggcctgc 4080

agctgcagaa ggtgactgtg ctggggggtgg ccactgcccc ccagcaggtg ctgagcaatg 4140

gggtgcctgt gagcaacttc acctatagcc ctgataccaa ggtgctggat atttgtgtga 4200

gcctgctgat gggggagcag ttcctggtga gctggtgctg actcgagaga tctaccggtg 4260

aattcaccgc gggtttaaac tgtgccttct agttgccagc catctgttgt ttgcccctcc 4320

cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 4380

gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtgggggct 4440

agctctagac tcgagatcca ctagggccgc aggaacccct agtgatggag ttggccactc 4500

cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 4560

gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag g 4611

<210> 22

<211> 4611

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 22

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggccgca ggctcagagg cacacaggag tttctgggct 180

caccctgccc ccttccaacc cctcagttcc catcctccag cagctgtttg tgtgctgcct 240

ctgaagtcca cactgaacaa acttcagcct actcatgtcc ctaaaatggg caaacattgc 300

aagcagcaaa cagcaaacac acagccctcc ctgcctgctg accttggagc tggggcagag 360

gtcagagacc tctctgggcc catgccacct ccaacatcca ctcgacccct tggaatttcg 420

gtggagagga gcagaggttg tcctggcgtg gtttaggtag tgtgagaggg gtacccgggg 480

atcttgctac cagtggaaca gccactaagg attctgcagt gagagcagag ggccagctaa 540

gtggtactct cccagagact gtctgactca cgccaccccc tccaccttgg acacaggacg 600

ctgtggtttc tgagccaggt acaatgactc ctttcggtaa gtgcagtgga agctgtacac 660

tgcccaggca aagcgtccgg gcagcgtagg cgggcgactc agatcccagc cagtggactt 720

agcccctgtt tgctcctccg ataactgggg tgaccttggt taatattcac cagcagcctc 780

ccccgttgcc cctctggatc cactgcttaa atacggacga ggacagggcc ctgtctcctc 840

agcttcaggc accaccactg acctgggaca gtgaatagat cctgagaact tcagggtgag 900

tctatgggac ccttgatgtt ttctttcccc ttcttttcta tggttaagtt catgtcatag 960

gaaggggaga agtaacaggg tacacatatt gaccaaatca gggtaatttt gcatttgtaa 1020

ttttaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt ctaatacttt 1080

ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct ttgcaccatt 1140

ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct gcatataaat 1200

atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata gcagctacaa 1260

tccagctacc attctgcttt tattttctgg ttgggataag gctggattat tctgagtcca 1320

agctaggccc ttttgctaat cttgttcata cctcttatct tcctcccaca gctcctgggc 1380

aacctgctgg tctctctgct ggcccatcac tttggcaaag cacgcgtgcc accatggctt 1440

tcctgtggct gctgtcttgt tgggctctgc tgggcaccac ctttggcctg ctggtgccca 1500

gggagctgtc tggcagcagc cctgtgctgg aggagaccca ccctgctcat cagcagggg 1560

ctagcaggcc tggccccagg gatgcccagg ctcaccctgg gagacccagg gctgtgccca 1620

ctcagtgtga tgtgcccccc aacagcaggt ttgactgtgc tcctgacaag gctatcaccc 1680

aggagcagtg tgaggccagg gggtgctgct acattcctgc taagcagggc ctgcaggggg 1740

cccagatggg ccagccctgg tgcttcttcc ccccctctta tcccagctat aagctggaga 1800

acctgagcag ctctgagatg ggctacactg ccaccctgac caggaccact cccaccttct 1860

ttcccaagga tattctgact ctgaggctgg atgtgatgat ggagactgag aacaggctgc 1920

acttcactat caaggaccct gccaatagga ggtatgaggt gcccctggag actcctcatg 1980

tgcatagcag ggccccttct cctctgtatt ctgtggagtt ctctgaggag ccctttgggg 2040

tgattgtgag gaggcagctg gatggcaggg tgctgctgaa caccactgtg gcccccctgt 2100

tctttgctga ccagttcctg cagctgagca cttctctgcc cagccagtac attactgggc 2160

tggctgagca tctgagcccc ctgatgctga gcacctcttg gaccaggatc accctgtgga 2220

acagggacct ggcccccact cctggggcta acctgtatgg ctctcacccc ttttacctgg 2280

ccctggagga tggggggctct gcccatgggg tgtttctgct gaacagcaat gctatggatg 2340

tggtgctgca gccctctcca gccctgtctt ggagggagcac tgggggcatt ctggatgtgt 2400

acattttcct ggggcctgaa cccaagtctg tggtgcagca gtacctggat gtggtgggct 2460

accccttcat gcccccctat tggggggctgg ggtttcacct gtgcaggtgg ggctacagca 2520

gcactgccat caccaggcag gtggtggaga acatgaccag ggcccatttc cccctggatg 2580

tgcagtggaa tgacctggac tacatggata gcaggaggga tttcaccttc aacaaggatg 2640

gcttcaggga ctttcctgcc atggtgcagg agctgcacca ggggggcagg aggtatatga 2700

tgattgtgga ccctgctatc agcagctctg gccctgctgg ctcttacagg ccctatgatg 2760

agggcctgag gaggggggtg tttatcacta atgaaactgg ccagcctctg attggcaagg 2820

tctggcctgg ctctactgcc ttccctgatt ttactaaccc cactgccctg gcctggtggg 2880

aggacatggt ggctgagttc catgatcagg tgccttttga tggcatgtgg attgatatga 2940

atgaaccaag caacttcatc agaggctctg aggatggctg ccccaacaat gagctggaga 3000

acccccccta tgtgcctggg gtggtggggg gcactctgca ggctgccacc atttgtgcta 3060

gcagccacca gttcctgagc acccactaca atctgcacaa cctgtatggc ctgactgaag 3120

ccattgccag ccatagggcc ctggtgaagg ccaggggcac taggcctttt gtgatcagca 3180

ggagcacttt tgctggccat ggcaggtatg ctggccactg gactggggat gtgtggagca 3240

gctgggagca gctggccagc tctgtgcctg agattctgca gtttaacctg ctgggggtgc 3300

ccctggtggg ggctgatgtg tgtggcttcc tgggcaacac ctctgaggag ctgtgtgtga 3360

ggtggaccca gctggggggcc ttttatccct tcatgaggaa ccacaacagc ctgctgagcc 3420

tgcctcagga gccctactct ttctctgagc ctgcccagca ggccatgagg aaggccctga 3480

ccctgaggta tgccctgctg ccccacctgt ataccctgtt ccaccaggcc catgtggctg 3540

gggagactgt ggccaggccc ctgttcctgg agttccccaa ggacagcagc acctggactg 3600

tggatcatca gctgctgtgg ggggaggccc tgctgatcac ccctgtgctg caggctggca 3660

aggctgaggt cactggctac ttccctctgg gcacctggta tgacctgcag actgtgcctg 3720

tggaggctct gggcagcctg cccccccccc ctgctgctcc cagggagcct gccatccact 3780

ctgaggcca gtgggtgacc ctgcctgctc ccctggacac catcaatgtg cacctgaggg 3840

ctggctacat tatccccctg cagggcccag ggctgactac cactgagagc agacagcagc 3900

ccatggctct ggctgtggcc ctgaccaagg ggggggaagc taggggggag ctgttctggg 3960

atgatgggga gagcctggag gtgctggaga ggggggccta tacccaggtg atcttcctgg 4020

ctaggaacaa caccattgtc aatgagctgg tgaggtgac ttctgagggg gctgggctgc 4080

agctgcagaa ggtgactgtg ctggggggtgg ccactgctcc ccagcaggtg ctgagcaatg 4140

gggtgcctgt gagcaacttc acctacagcc ctgacaccaa ggtgctggac atctgtgtgt 4200

ctctgctgat ggggggagcag ttcctggtga gctggtgctg actcgagaga tctaccggtg 4260

aattcaccgc gggtttaaac tgtgccttct agttgccagc catctgttgt ttgcccctcc 4320

cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 4380

gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtgggggct 4440

agctctagac tcgagatcca ctagggccgc aggaacccct agtgatggag ttggccactc 4500

cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 4560

gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag g 4611

<210> 23

<211> 4611

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 23

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggccgca ggctcagagg cacacaggag tttctgggct 180

caccctgccc ccttccaacc cctcagttcc catcctccag cagctgtttg tgtgctgcct 240

ctgaagtcca cactgaacaa acttcagcct actcatgtcc ctaaaatggg caaacattgc 300

aagcagcaaa cagcaaacac acagccctcc ctgcctgctg accttggagc tggggcagag 360

gtcagagacc tctctgggcc catgccacct ccaacatcca ctcgacccct tggaatttcg 420

gtggagagga gcagaggttg tcctggcgtg gtttaggtag tgtgagaggg gtacccgggg 480

atcttgctac cagtggaaca gccactaagg attctgcagt gagagcagag ggccagctaa 540

gtggtactct cccagagact gtctgactca cgccaccccc tccaccttgg acacaggacg 600

ctgtggtttc tgagccaggt acaatgactc ctttcggtaa gtgcagtgga agctgtacac 660

tgcccaggca aagcgtccgg gcagcgtagg cgggcgactc agatcccagc cagtggactt 720

agcccctgtt tgctcctccg ataactgggg tgaccttggt taatattcac cagcagcctc 780

ccccgttgcc cctctggatc cactgcttaa atacggacga ggacagggcc ctgtctcctc 840

agcttcaggc accaccactg acctgggaca gtgaatagat cctgagaact tcagggtgag 900

tctatgggac ccttgatgtt ttctttcccc ttcttttcta tggttaagtt catgtcatag 960

gaaggggaga agtaacaggg tacacatatt gaccaaatca gggtaatttt gcatttgtaa 1020

ttttaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt ctaatacttt 1080

ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct ttgcaccatt 1140

ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct gcatataaat 1200

atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata gcagctacaa 1260

tccagctacc attctgcttt tattttctgg ttgggataag gctggattat tctgagtcca 1320

agctaggccc ttttgctaat cttgttcata cctcttatct tcctcccaca gctcctgggc 1380

aacctgctgg tctctctgct ggcccatcac tttggcaaag cacgcgtgcc accatggcct 1440

tcctgtggct gctgtcttgc tgggctctgc tggggaccac ctttggcctg ctggtcccca 1500

gggagctgtc tggctcttct cctgtcctgg aggagaccca ccctgcccac cagcagggg 1560

ctagcaggcc tggccccagg gatgcccagg cccaccctgg caggcccagg gctgtgccca 1620

cccagtgtga tgtgcctccc aacagcaggt ttgactgtgc ccctgacaag gccatcaccc 1680

aggagcagtg tgaggccagg ggctgctgct atatccctgc caagcagggc ctgcaggggg 1740

ctcagatggg ccagccctgg tgcttctttc ccccctctta tcctagctat aagctggaga 1800

acctgagcag ctctgagatg gggtacactg ccaccctgac caggaccacc cccactttct 1860

tccctaagga catcctgacc ctgaggctgg atgtgatgat ggagactgag aataggctgc 1920

actttactat caaggaccct gccaacagga ggtatgaggt gcctctggag accccccatg 1980

tgcattctag ggcccccagc cccctgtact ctgtggagtt ctctgaggag ccctttgggg 2040

tgattgtgag gagacagctg gatggcaggg tcctgctgaa caccactgtg gctcccctgt 2100

tttttgctga ccagttcctg cagctgagca ccagcctgcc cagccagtac atcactgggc 2160

tggctgagca cctgagcccc ctgatgctga gcaccagctg gaccaggatc accctgtgga 2220

acagggatct ggctcctacc cctggggcca acctgtatgg ctctcacccc ttttacctgg 2280

ccctggagga tggggggctct gcccatgggg tgttcctgct gaacagcaat gctatggatg 2340

tggtgctgca gcccagccct gccctgagct ggaggtctac tgggggcatc ctggatgtgt 2400

acatctttct ggggcctgag cccaagtctg tggtgcagca gtacctggat gtggtgggct 2460

atccttttat gcccccctat tggggcctgg gcttccacct gtgcaggtgg ggctacagca 2520

gcactgccat caccagacag gtggtggaga acatgaccag ggcccacttc cccctggatg 2580

tgcagtggaa tgacctggac tacatggaca gcaggaggga cttcaccttt aacaaggatg 2640

gctttaggga cttccctgcc atggtgcagg agctgcatca ggggggcagg aggtacatga 2700

tgattgtgga cccagccatc agcagctctg ggcctgctgg gtcttacagg ccctatgatg 2760

agggcctgag gaggggggtg ttcatcacca atgagactgg ccagcccctg attggcaagg 2820

tgtggcctgg gagcactgcc ttccctgatt ttaccaaccc cactgccctg gcctggtggg 2880

aggatatggt ggctgagttt catgaccagg tgccctttga tggcatgtgg attgacatga 2940

atgagcccag caatttcatc aggggctctg aggatggctg ccccaacaat gagctggaga 3000

atcctcccta tgtgcctggg gtggtggggg gcaccctgca ggctgccacc atctgtgcct 3060

ctagccacca gttcctgagc acccactata acctgcataa cctgtatggc ctgactgagg 3120

ccattgccag ccatagagcc ctggtgaagg ccagagggac caggcccttt gtgatctcta 3180

ggagcacctt tgctggccat ggcaggtatg ctggccactg gactggggat gtgtggagct 3240

cttgggagca gctggccagc tctgtgccag agatcctgca gttcaacctg ctggggggtgc 3300

ctctggtggg ggctgatgtg tgtggcttcc tgggcaatac ctctgaagag ctgtgtgtga 3360

ggtggactca gctggggggcc ttctatccct tcatgaggaa ccacaacagc ctgctgtctc 3420

tgccccagga gccctacagc ttctctgagc ctgctcagca ggctatgagg aaggccctga 3480

ccctgaggta tgccctgctg ccccatctgt acaccctgtt ccaccaggcc catgtggctg 3540

gggagactgt ggccaggccc ctgtttctgg agtttcccaa ggacagcagc acctggactg 3600

tggaccatca gctgctgtgg ggggaggctc tgctgattac ccctgtgctg caggctggca 3660

aggctgaggt gactgggtac ttccccctgg ggacttggta tgacctgcag actgtgcctg 3720

tggaagctct gggcagcctg cccccacccc ctgctgcccc tagggagcct gccatccact 3780

ctgaggcca gtgggtgacc ctgcctgccc ctctggacac catcaatgtg cacctgaggg 3840

ctggctatat catccccctg cagggccctg ggctgaccac cactgagagc aggcagcagc 3900

ccatggccct ggctgtggcc ctgactaagg ggggggaggc caggggggag ctgttctggg 3960

atgatgggga gagcctggag gtgctggaga gaggggccta cacccaggtg atctttctgg 4020

caggaacaa caccattgtg aatgagctgg tgaggtgac ttctgagggg gctggcctgc 4080

agctgcagaa ggtgactgtg ctggggggtgg ccactgcccc ccagcaggtg ctgagcaatg 4140

gggtgcctgt gtctaacttc acctacagcc ctgatactaa ggtgctggat atctgtgtga 4200

gcctgctgat gggggagcag tttctggtga gctggtgctg actcgagaga tctaccggtg 4260

aattcaccgc gggtttaaac tgtgccttct agttgccagc catctgttgt ttgcccctcc 4320

cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 4380

gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtgggggct 4440

agctctagac tcgagatcca ctagggccgc aggaacccct agtgatggag ttggccactc 4500

cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 4560

gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag g 4611

<210> 24

<211> 4611

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 24

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggccgca ggctcagagg cacacaggag tttctgggct 180

caccctgccc ccttccaacc cctcagttcc catcctccag cagctgtttg tgtgctgcct 240

ctgaagtcca cactgaacaa acttcagcct actcatgtcc ctaaaatggg caaacattgc 300

aagcagcaaa cagcaaacac acagccctcc ctgcctgctg accttggagc tggggcagag 360

gtcagagacc tctctgggcc catgccacct ccaacatcca ctcgacccct tggaatttcg 420

gtggagagga gcagaggttg tcctggcgtg gtttaggtag tgtgagaggg gtacccgggg 480

atcttgctac cagtggaaca gccactaagg attctgcagt gagagcagag ggccagctaa 540

gtggtactct cccagagact gtctgactca cgccaccccc tccaccttgg acacaggacg 600

ctgtggtttc tgagccaggt acaatgactc ctttcggtaa gtgcagtgga agctgtacac 660

tgcccaggca aagcgtccgg gcagcgtagg cgggcgactc agatcccagc cagtggactt 720

agcccctgtt tgctcctccg ataactgggg tgaccttggt taatattcac cagcagcctc 780

ccccgttgcc cctctggatc cactgcttaa atacggacga ggacagggcc ctgtctcctc 840

agcttcaggc accaccactg acctgggaca gtgaatagat cctgagaact tcagggtgag 900

tctatgggac ccttgatgtt ttctttcccc ttcttttcta tggttaagtt catgtcatag 960

gaaggggaga agtaacaggg tacacatatt gaccaaatca gggtaatttt gcatttgtaa 1020

ttttaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt ctaatacttt 1080

ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct ttgcaccatt 1140

ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct gcatataaat 1200

atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata gcagctacaa 1260

tccagctacc attctgcttt tattttctgg ttgggataag gctggattat tctgagtcca 1320

agctaggccc ttttgctaat cttgttcata cctcttatct tcctcccaca gctcctgggc 1380

aacctgctgg tctctctgct ggcccatcac tttggcaaag cacgcgtgcc accatggcct 1440

ttctgtggct gctgtcctgc tgggccctgc tggggaccac ctttggcctg ctggtgccca 1500

gggagctgtc tgggagcagc ccagtgctgg aggagaccca ccctgcccac cagcaggggg 1560

ccagcaggcc tggccctagg gatgcccagg cccaccctgg caggcccagg gctgtgccta 1620

cccagtgtga tgtgccaccc aattctaggt ttgactgtgc tcctgacaag gccatcactc 1680

aggagcagtg tgaagctagg gggtgctgct acatcccagc caagcagggc ctgcaggggg 1740

cccagatggg ccagccctgg tgcttcttcc cccccagcta ccctagctac aagctggaga 1800

atctgagcag ctctgagatg ggctacactg ctaccctgac caggaccact cctaccttct 1860

tccccaagga catcctgact ctgaggctgg atgtcatgat ggagactgaa aataggctgc 1920

acttcaccat caaggaccct gccaatagga ggtatgaggt gcctctggag accccccatg 1980

tgcatagcag ggctcccagc cccctgtatt ctgtggagtt ctctgaggag ccctttgggg 2040

tcattgtgag gagacagctg gatgggagg tgctgctgaa cactactgtg gctcccctgt 2100

tctttgctga ccagttcctg cagctgtcta ccagcctgcc cagccagtac atcactgggc 2160

tggctgagca tctgagcccc ctgatgctga gcaccagctg gaccaggatc actctgtgga 2220

acagggatct ggcccccact cctggggcca acctgtatgg gagccatccc ttctacctgg 2280

ccctggagga tggggggctct gcccatgggg tgttcctgct gaacagcaat gccatggatg 2340

tggtgctgca gcctagccct gccctgagct ggagaggcac tgggggcatc ctggatgtct 2400

acatcttcct ggggcctgag cccaagtctg tggtgcagca gtatctggat gtggtggggt 2460

atcccttcat gcccccctac tggggcctgg gctttcacct gtgcaggtgg ggctacagca 2520

gcactgccat caccaggcag gtggtggaga acatgaccag ggcccacttc cctctggatg 2580

tgcagtggaa tgacctggac tatatggatt ctaggagaga ctttactttt aacaaggatg 2640

gcttcaggga tttccctgcc atggtgcagg agctgcacca ggggggcagg aggtacatga 2700

tgattgtgga ccctgctatt agcagctctg gccctgctgg gtcttacagg ccttatgatg 2760

agggcctgag gaggggggtg ttcatcacca atgagactgg ccagcccctg attggcaagg 2820

tgtggcctgg cagcactgcc ttccctgact tcaccaaccc cactgccctg gcctggtggg 2880

aggacatggt ggctgagttc catgaccagg tgccctttga tgggatgtgg attgacatga 2940

atgagccctc taacttcatc agggggtctg aggatggctg ccccaacaat gagctggaga 3000

acccccccta tgtgcctggg gtggtggggg gcactctgca ggctgccact atctgtgctt 3060

cttctcacca gtttctgagc acccactata atctgcacaa cctgtatggc ctgactgagg 3120

ccattgccag ccatagggcc ctggtgaagg ccaggggcac caggcccttt gtgatcagca 3180

ggtctacctt tgctggccat ggcaggtatg ctggccactg gactggggat gtgtggtctt 3240

cttgggagca gctggccagc tctgtgcctg agatcctgca gttcaacctg ctggggggtgc 3300

ctctggtggg ggctgatgtg tgtggctttc tgggcaacac ctctgaggag ctgtgtgtga 3360

ggtggaccca gctggggggcc ttttacccct tcatgaggaa ccacaatagc ctgctgagcc 3420

tgccccagga gccttactct ttctctgagc ctgcccagca ggccatgagg aaggccctga 3480

ctctgaggta tgccctgctg ccccatctgt ataccctgtt tcaccaggcc catgtggctg 3540

gggagactgt ggctaggcct ctgtttctgg agttccctaa ggactctagc acctggactg 3600

tggaccacca gctgctgtgg ggggaggccc tgctgatcac ccctgtgctg caggctggca 3660

aggctgaggt gactggctac ttccccctgg gcacctggta tgacctgcag actgtgcctg 3720

tggaggccct ggggagcctg cctccccccc ctgctgcccc cagggagcct gccattcatt 3780

ctgaggcca gtgggtgacc ctgcctgccc ctctggacac catcaatgtg cacctgaggg 3840

ctgggtacat catccccctg cagggccctg gcctgaccac cactgagagc aggcagcagc 3900

ccatggccct ggctgtggct ctgaccaagg ggggggaggc caggggggag ctgttctggg 3960

atgatgggga gtctctggag gtgctggaga ggggggccta cacccaggtg atctttctgg 4020

caggaacaa tactattgtg aatgagctgg tgaggtgac ctctgagggg gctggcctgc 4080

agctgcagaa ggtgactgtg ctggggggtgg ccactgcccc ccagcaggtc ctgagcaatg 4140

gggtgcctgt gagcaacttc acctactctc ctgacaccaa ggtgctggac atttgtgtgt 4200

ctctgctgat ggggggagcag ttcctggtga gctggtgctg actcgagaga tctaccggtg 4260

aattcaccgc gggtttaaac tgtgccttct agttgccagc catctgttgt ttgcccctcc 4320

cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 4380

gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtgggggct 4440

agctctagac tcgagatcca ctagggccgc aggaacccct agtgatggag ttggccactc 4500

cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 4560

gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag g 4611

<210> 25

<211> 935

<212> PROTEIN

<213> Artificial sequence

<220>

<223> GAA peptide

<400> 25

Met Ala Phe Leu Trp Leu Leu Ser Cys Trp Ala Leu Leu Gly Thr Thr

1 5 10 15

Phe Gly Leu Leu Val Pro Arg Glu Leu Ser Gly Ser Ser Pro Val Leu

20 25 30

Glu Glu Thr His Pro Ala His Gln Gln Gly Ala Ser Arg Pro Gly Pro

35 40 45

Arg Asp Ala Gln Ala His Pro Gly Arg Pro Arg Ala Val Pro Thr Gln

50 55 60

Cys Asp Val Pro Pro Asn Ser Arg Phe Asp Cys Ala Pro Asp Lys Ala

65 70 75 80

Ile Thr Gln Glu Gln Cys Glu Ala Arg Gly Cys Cys Tyr Ile Pro Ala

85 90 95

Lys Gln Gly Leu Gln Gly Ala Gln Met Gly Gln Pro Trp Cys Phe Phe

100 105 110

Pro Pro Ser Tyr Pro Ser Tyr Lys Leu Glu Asn Leu Ser Ser Ser Glu

115 120 125

Met Gly Tyr Thr Ala Thr Leu Thr Arg Thr Thr Pro Thr Phe Phe Pro

130 135 140

Lys Asp Ile Leu Thr Leu Arg Leu Asp Val Met Met Glu Thr Glu Asn

145 150 155 160

Arg Leu His Phe Thr Ile Lys Asp Pro Ala Asn Arg Arg Tyr Glu Val

165 170 175

Pro Leu Glu Thr Pro His Val His Ser Arg Ala Pro Ser Pro Leu Tyr

180 185 190

Ser Val Glu Phe Ser Glu Glu Pro Phe Gly Val Ile Val Arg Arg Gln

195 200 205

Leu Asp Gly Arg Val Leu Leu Asn Thr Thr Val Ala Pro Leu Phe Phe

210 215 220

Ala Asp Gln Phe Leu Gln Leu Ser Thr Ser Leu Pro Ser Gln Tyr Ile

225 230 235 240

Thr Gly Leu Ala Glu His Leu Ser Pro Leu Met Leu Ser Thr Ser Trp

245 250 255

Thr Arg Ile Thr Leu Trp Asn Arg Asp Leu Ala Pro Thr Pro Gly Ala

260 265 270

Asn Leu Tyr Gly Ser His Pro Phe Tyr Leu Ala Leu Glu Asp Gly Gly

275 280 285

Ser Ala His Gly Val Phe Leu Leu Asn Ser Asn Ala Met Asp Val Val

290 295 300

Leu Gln Pro Ser Pro Ala Leu Ser Trp Arg Ser Thr Gly Gly Ile Leu

305 310 315 320

Asp Val Tyr Ile Phe Leu Gly Pro Glu Pro Lys Ser Val Val Gln Gln

325 330 335

Tyr Leu Asp Val Val Gly Tyr Pro Phe Met Pro Pro Tyr Trp Gly Leu

340 345 350

Gly Phe His Leu Cys Arg Trp Gly Tyr Ser Ser Thr Ala Ile Thr Arg

355 360 365

Gln Val Val Glu Asn Met Thr Arg Ala His Phe Pro Leu Asp Val Gln

370 375 380

Trp Asn Asp Leu Asp Tyr Met Asp Ser Arg Arg Asp Phe Thr Phe Asn

385 390 395 400

Lys Asp Gly Phe Arg Asp Phe Pro Ala Met Val Gln Glu Leu His Gln

405 410 415

Gly Gly Arg Arg Tyr Met Met Ile Val Asp Pro Ala Ile Ser Ser Ser

420 425 430

Gly Pro Ala Gly Ser Tyr Arg Pro Tyr Asp Glu Gly Leu Arg Arg Gly

435 440 445

Val Phe Ile Thr Asn Glu Thr Gly Gln Pro Leu Ile Gly Lys Val Trp

450 455 460

Pro Gly Ser Thr Ala Phe Pro Asp Phe Thr Asn Pro Thr Ala Leu Ala

465 470 475 480

Trp Trp Glu Asp Met Val Ala Glu Phe His Asp Gln Val Pro Phe Asp

485 490 495

Gly Met Trp Ile Asp Met Asn Glu Pro Ser Asn Phe Ile Arg Gly Ser

500 505 510

Glu Asp Gly Cys Pro Asn Asn Glu Leu Glu Asn Pro Pro Tyr Val Pro

515 520 525

Gly Val Val Gly Gly Thr Leu Gln Ala Ala Thr Ile Cys Ala Ser Ser

530 535 540

His Gln Phe Leu Ser Thr His Tyr Asn Leu His Asn Leu Tyr Gly Leu

545 550 555 560

Thr Glu Ala Ile Ala Ser His Arg Ala Leu Val Lys Ala Arg Gly Thr

565 570 575

Arg Pro Phe Val Ile Ser Arg Ser Thr Phe Ala Gly His Gly Arg Tyr

580 585 590

Ala Gly His Trp Thr Gly Asp Val Trp Ser Ser Trp Glu Gln Leu Ala

595 600 605

Ser Ser Val Pro Glu Ile Leu Gln Phe Asn Leu Leu Gly Val Pro Leu

610 615 620

Val Gly Ala Asp Val Cys Gly Phe Leu Gly Asn Thr Ser Glu Glu Leu

625 630 635 640

Cys Val Arg Trp Thr Gln Leu Gly Ala Phe Tyr Pro Phe Met Arg Asn

645 650 655

His Asn Ser Leu Leu Ser Leu Pro Gln Glu Pro Tyr Ser Phe Ser Glu

660 665 670

Pro Ala Gln Gln Ala Met Arg Lys Ala Leu Thr Leu Arg Tyr Ala Leu

675 680 685

Leu Pro His Leu Tyr Thr Leu Phe His Gln Ala His Val Ala Gly Glu

690 695 700

Thr Val Ala Arg Pro Leu Phe Leu Glu Phe Pro Lys Asp Ser Ser Thr

705 710 715 720

Trp Thr Val Asp His Gln Leu Leu Trp Gly Glu Ala Leu Leu Ile Thr

725 730 735

Pro Val Leu Gln Ala Gly Lys Ala Glu Val Thr Gly Tyr Phe Pro Leu

740 745 750

Gly Thr Trp Tyr Asp Leu Gln Thr Val Pro Val Glu Ala Leu Gly Ser

755 760 765

Leu Pro Pro Pro Pro Ala Ala Pro Arg Glu Pro Ala Ile His Ser Glu

770 775 780

Gly Gln Trp Val Thr Leu Pro Ala Pro Leu Asp Thr Ile Asn Val His

785 790 795 800

Leu Arg Ala Gly Tyr Ile Ile Pro Leu Gln Gly Pro Gly Leu Thr Thr

805 810 815

Thr Glu Ser Arg Gln Gln Pro Met Ala Leu Ala Val Ala Leu Thr Lys

820 825 830

Gly Gly Glu Ala Arg Gly Glu Leu Phe Trp Asp Asp Gly Glu Ser Leu

835 840 845

Glu Val Leu Glu Arg Gly Ala Tyr Thr Gln Val Ile Phe Leu Ala Arg

850 855 860

Asn Asn Thr Ile Val Asn Glu Leu Val Arg Val Thr Ser Glu Gly Ala

865 870 875 880

Gly Leu Gln Leu Gln Lys Val Thr Val Leu Gly Val Ala Thr Ala Pro

885 890 895

Gln Gln Val Leu Ser Asn Gly Val Pro Val Ser Asn Phe Thr Tyr Ser

900 905 910

Pro Asp Thr Lys Val Leu Asp Ile Cys Val Ser Leu Leu Met Gly Glu

915 920 925

Gln Phe Leu Val Ser Trp Cys

930 935

<210> 26

<211> 268

<212> DNA

<213> Artificial sequence

<220>

<223> Modified nucleic acid

<400> 26

agatctagag ctgaattcct gcagccaggg ggatcagcct ctactgtgcc ttctagttgc 60

cagccatctg ttgtttgccc ctcccccttg ccttccttga ccctggaagg tgccactccc 120

actgtccttt cctaataaaa tgaggaaatt gcatcacatt gtctgagtag gtgtcattct 180

attctgggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 240

catgctgggg atgcagtggg ctctatgg 268

<210> 27

<211> 202

<212> DNA

<213> Bos taurus

<400> 27

agatctaccg gtgaattcac cgcgggttta aactgtgcct tctagttgcc agccatctgt 60

tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 120

ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 180

tggggtgggg gctagctcta ga 202

<210> 28

<211> 456

<212> DNA

<213> Homo sapiens

<400> 28

gggcccatgc cacctccaac atccactcga ccccttggaa tttcggtgga gaggagcaga 60

ggttgtcctg gcgtggttta ggtagtgtga gaggggtacc cggggatctt gctaccagtg 120

gaacagccac taaggattct gcagtgagag cagagggcca gctaagtggt actctcccag 180

agactgtctg actcacgcca ccccctccac cttggacaca ggacgctgtg gtttctgagc 240

caggtacaat gactcctttc ggtaagtgca gtggaagctg tacactgccc aggcaaagcg 300

tccgggcagc gtaggcgggc gactcagatc ccagccagtg gacttagccc ctgtttgctc 360

ctccgataac tggggtgacc ttggttaata ttcaccagca gcctcccccg ttgcccctct 420

ggatccactg cttaaatacg gacgaggaca gggccc 456

<210> 29

<211> 444

<212> DNA

<213> Homo sapiens

<400> 29

atgccacctc caacatccac tcgacccctt ggaatttcgg tggagaggag cagaggttgt 60

cctggcgtgg tttaggtagt gtgagagggg tacccgggga tcttgctacc agtggaacag 120

ccactaagga ttctgcagtg agagcagagg gccagctaag tggtactctc ccagagactg 180

tctgactcac gccaccccct ccaccttgga cacaggacgc tgtggtttct gagccaggta 240

caatgactcc tttcggtaag tgcagtggaa gctgtacact gcccaggcaa agcgtccggg 300

cagcgtaggc gggcgactca gatcccagcc agtggactta gcccctgttt gctcctccga 360

taactggggt gaccttggtt aatattcacc agcagcctcc cccgttgccc ctctggatcc 420

actgcttaaa tacggacgag gaca 444

<210> 30

<211> 738

<212> PROTEIN

<213> Artificial sequence

<220>

<223> AAV vector capsid

<400> 30

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro

20 25 30

Lys Ala Asn Gln Gln Lys Gln Asp Asn Gly Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Gln Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Ser Pro Val Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Pro Ser Pro Gln Arg Ser Pro Asp Ser Ser Thr Gly Ile

145 150 155 160

Gly Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln

165 170 175

Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro

180 185 190

Pro Ala Ala Pro Ser Gly Val Gly Pro Asn Thr Met Ala Ala Gly Gly

195 200 205

Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser

210 215 220

Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val

225 230 235 240

Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His

245 250 255

Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly Gly Ser Thr Asn Asp

260 265 270

Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn

275 280 285

Arg Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn

290 295 300

Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn

305 310 315 320

Ile Gln Val Lys Glu Val Thr Gln Asn Glu Gly Thr Lys Thr Ile Ala

325 330 335

Asn Asn Leu Thr Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln

340 345 350

Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe

355 360 365

Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn

370 375 380

Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr

385 390 395 400

Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr

405 410 415

Asn Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser

420 425 430

Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu

435 440 445

Ser Arg Thr Gln Ser Thr Gly Gly Thr Ala Gly Thr Gln Gln Leu Leu

450 455 460

Phe Ser Gln Ala Gly Pro Asn Asn Met Ser Ala Gln Ala Lys Asn Trp

465 470 475 480

Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Thr Thr Leu Ser

485 490 495

Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His

500 505 510

Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr

515 520 525

His Lys Asp Asp Glu Glu Arg Phe Phe Pro Ser Ser Gly Val Leu Met

530 535 540

Phe Gly Lys Gln Gly Ala Gly Lys Asp Asn Val Asp Tyr Ser Ser Val

545 550 555 560

Met Leu Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr

565 570 575

Glu Gln Tyr Gly Val Val Ala Asp Asn Leu Gln Gln Gln Asn Ala Ala

580 585 590

Pro Ile Val Gly Ala Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val

595 600 605

Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile

610 615 620

Pro His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe

625 630 635 640

Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val

645 650 655

Pro Ala Asp Pro Pro Thr Thr Phe Asn Gln Ala Lys Leu Ala Ser Phe

660 665 670

Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu

675 680 685

Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr

690 695 700

Ser Asn Tyr Tyr Lys Ser Thr Asn Val Asp Phe Ala Val Asn Thr Glu

705 710 715 720

Gly Thr Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg

725 730 735

Asn Leu

<210> 31

<211> 601

<212> PROTEIN

<213> Artificial sequence

<220>

<223> AAV vector capsid

<400> 31

Thr Ala Pro Gly Lys Lys Arg Pro Val Glu Pro Ser Pro Gln Arg Ser

1 5 10 15

Pro Asp Ser Ser Thr Gly Ile Gly Lys Lys Gly Gln Gln Pro Ala Lys

20 25 30

Lys Arg Leu Asn Phe Gly Gln Thr Gly Asp Ser Glu Ser Val Pro Asp

35 40 45

Pro Gln Pro Ile Gly Glu Pro Pro Ala Gly Pro Ser Gly Leu Gly Ser

50 55 60

Gly Thr Met Ala Ala Gly Gly Gly Ala Pro Met Ala Asp Asn Asn Glu

65 70 75 80

Gly Ala Asp Gly Val Gly Ser Ser Ser Gly Asn Trp His Cys Asp Ser

85 90 95

Thr Trp Leu Gly Asp Arg Val Ile Thr Thr Ser Thr Arg Thr Trp Ala

100 105 110

Leu Pro Thr Tyr Asn Asn His Leu Tyr Lys Gln Ile Ser Asn Gly Thr

115 120 125

Ser Gly Gly Ser Thr Asn Asp Asn Thr Tyr Phe Gly Tyr Ser Thr Pro

130 135 140

Trp Gly Tyr Phe Asp Phe Asn Arg Phe His Cys His Phe Ser Pro Arg

145 150 155 160

Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp Gly Phe Arg Pro Lys Arg

165 170 175

Leu Asn Phe Lys Leu Phe Asn Ile Gln Val Lys Glu Val Thr Gln Asn

180 185 190

Glu Gly Thr Lys Thr Ile Ala Asn Asn Leu Thr Ser Thr Ile Gln Val

195 200 205

Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr Val Leu Gly Ser Ala His

210 215 220

Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp Val Phe Met Ile Pro Gln

225 230 235 240

Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser Gln Ala Val Gly Arg Ser

245 250 255

Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser Gln Met Leu Arg Thr Gly

260 265 270

Asn Asn Phe Glu Phe Ser Tyr Asn Phe Glu Asp Val Pro Phe His Ser

275 280 285

Ser Tyr Ala His Ser Gln Ser Leu Asp Arg Leu Met Asn Pro Leu Ile

290 295 300

Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg Thr Gln Ser Thr Gly Gly Thr

305 310 315 320

Ala Gly Thr Gln Gln Leu Leu Phe Ser Gln Ala Gly Pro Asn Asn Met

325 330 335

Ser Ala Gln Ala Lys Asn Trp Leu Pro Gly Pro Cys Tyr Arg Gln Gln

340 345 350

Arg Val Ser Thr Thr Leu Ser Gln Asn Asn Asn Ser Asn Phe Ala Trp

355 360 365

Thr Gly Ala Thr Lys Tyr His Leu Asn Gly Arg Asp Ser Leu Val Asn

370 375 380

Pro Gly Val Ala Met Ala Thr His Lys Asp Asp Glu Glu Arg Phe Phe

385 390 395 400

Pro Ser Ser Gly Val Leu Met Phe Gly Lys Gln Gly Ala Gly Lys Asp

405 410 415

Asn Val Asp Tyr Ser Ser Val Met Leu Thr Ser Glu Glu Glu Ile Lys

420 425 430

Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr Gly Val Val Ala Asp Asn

435 440 445

Leu Gln Gln Gln Asn Ala Ala Pro Ile Val Gly Ala Val Asn Ser Gln

450 455 460

Gly Ala Leu Pro Gly Met Val Trp Gln Asn Arg Asp Val Tyr Leu Gln

465 470 475 480

Gly Pro Ile Trp Ala Lys Ile Pro His Thr Asp Gly Asn Phe His Pro

485 490 495

Ser Pro Leu Met Gly Gly Phe Gly Leu Lys His Pro Pro Pro Gln Ile

500 505 510

Leu Ile Lys Asn Thr Pro Val Pro Ala Asp Pro Pro Thr Thr Phe Asn

515 520 525

Gln Ala Lys Leu Ala Ser Phe Ile Thr Gln Tyr Ser Thr Gly Gln Val

530 535 540

Ser Val Glu Ile Glu Trp Glu Leu Gln Lys Glu Asn Ser Lys Arg Trp

545 550 555 560

Asn Pro Glu Ile Gln Tyr Thr Ser Asn Tyr Tyr Lys Ser Thr Asn Val

565 570 575

Asp Phe Ala Val Asn Thr Glu Gly Thr Tyr Ser Glu Pro Arg Pro Ile

580 585 590

Gly Thr Arg Tyr Leu Thr Arg Asn Leu

595 600

<210> 32

<211> 535

<212> PROTEIN

<213> Artificial sequence

<220>

<223> AAV vector capsid

<400> 32

Met Ala Ala Gly Gly Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala

1 5 10 15

Asp Gly Val Gly Ser Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp

20 25 30

Leu Gly Asp Arg Val Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro

35 40 45

Thr Tyr Asn Asn His Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly

50 55 60

Gly Ser Thr Asn Asp Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly

65 70 75 80

Tyr Phe Asp Phe Asn Arg Phe His Cys His Phe Ser Pro Arg Asp Trp

85 90 95

Gln Arg Leu Ile Asn Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn

100 105 110

Phe Lys Leu Phe Asn Ile Gln Val Lys Glu Val Thr Gln Asn Glu Gly

115 120 125

Thr Lys Thr Ile Ala Asn Asn Leu Thr Ser Thr Ile Gln Val Phe Thr

130 135 140

Asp Ser Glu Tyr Gln Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly

145 150 155 160

Cys Leu Pro Pro Phe Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly

165 170 175

Tyr Leu Thr Leu Asn Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe

180 185 190

Tyr Cys Leu Glu Tyr Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn

195 200 205

Phe Glu Phe Ser Tyr Asn Phe Glu Asp Val Pro Phe His Ser Ser Tyr

210 215 220

Ala His Ser Gln Ser Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln

225 230 235 240

Tyr Leu Tyr Tyr Leu Ser Arg Thr Gln Ser Thr Gly Gly Thr Ala Gly

245 250 255

Thr Gln Gln Leu Leu Phe Ser Gln Ala Gly Pro Asn Asn Met Ser Ala

260 265 270

Gln Ala Lys Asn Trp Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val

275 280 285

Ser Thr Thr Leu Ser Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly

290 295 300

Ala Thr Lys Tyr His Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly

305 310 315 320

Val Ala Met Ala Thr His Lys Asp Asp Glu Glu Arg Phe Phe Pro Ser

325 330 335

Ser Gly Val Leu Met Phe Gly Lys Gln Gly Ala Gly Lys Asp Asn Val

340 345 350

Asp Tyr Ser Ser Val Met Leu Thr Ser Glu Glu Glu Ile Lys Thr Thr

355 360 365

Asn Pro Val Ala Thr Glu Gln Tyr Gly Val Val Ala Asp Asn Leu Gln

370 375 380

Gln Gln Asn Ala Ala Pro Ile Val Gly Ala Val Asn Ser Gln Gly Ala

385 390 395 400

Leu Pro Gly Met Val Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro

405 410 415

Ile Trp Ala Lys Ile Pro His Thr Asp Gly Asn Phe His Pro Ser Pro

420 425 430

Leu Met Gly Gly Phe Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile

435 440 445

Lys Asn Thr Pro Val Pro Ala Asp Pro Pro Thr Thr Phe Asn Gln Ala

450 455 460

Lys Leu Ala Ser Phe Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val

465 470 475 480

Glu Ile Glu Trp Glu Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro

485 490 495

Glu Ile Gln Tyr Thr Ser Asn Tyr Tyr Lys Ser Thr Asn Val Asp Phe

500 505 510

Ala Val Asn Thr Glu Gly Thr Tyr Ser Glu Pro Arg Pro Ile Gly Thr

515 520 525

Arg Tyr Leu Thr Arg Asn Leu

530 535

<210> 33

<211> 736

<212> PROTEIN

<213> Artificial sequence

<220>

<223> AAV vector capsid

<400> 33

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Gln Pro Gly Ala Pro Lys Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Asp Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Val Gly

145 150 155 160

Lys Ser Gly Lys Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro

180 185 190

Ala Ala Pro Thr Ser Leu Gly Ser Asn Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Asn Asp Asn His Tyr

260 265 270

Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe His

275 280 285

Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp

290 295 300

Gly Phe Arg Pro Lys Lys Leu Ser Phe Lys Leu Phe Asn Ile Gln Val

305 310 315 320

Lys Glu Val Thr Gln Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn Leu

325 330 335

Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr

340 345 350

Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp

355 360 365

Val Phe Met Val Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser

370 375 380

Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser

385 390 395 400

Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Thr Phe Glu

405 410 415

Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg

420 425 430

Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Asn Arg Thr

435 440 445

Gln Gly Thr Thr Ser Gly Thr Thr Asn Gln Ser Arg Leu Leu Phe Ser

450 455 460

Gln Ala Gly Pro Gln Ser Met Ser Leu Gln Ala Arg Asn Trp Leu Pro

465 470 475 480

Gly Pro Cys Tyr Arg Gln Gln Arg Leu Ser Lys Thr Ala Asn Asp Asn

485 490 495

Asn Asn Ser Asn Phe Pro Trp Thr Ala Ala Ser Lys Tyr His Leu Asn

500 505 510

Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Asp Asp Glu Glu Lys Phe Phe Pro Met His Gly Asn Leu Ile Phe Gly

530 535 540

Lys Glu Gly Thr Thr Ala Ser Asn Ala Glu Leu Asp Asn Val Met Ile

545 550 555 560

Thr Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln

565 570 575

Tyr Gly Thr Val Ala Asn Asn Leu Gln Ser Ser Asn Thr Ala Pro Thr

580 585 590

Thr Arg Thr Val Asn Asp Gln Gly Ala Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu

625 630 635 640

Lys His Pro Pro Pro Gln Ile Met Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Asn Lys Ser Val Asn Val Asp Phe Thr Val Asp Thr Asn Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Pro Leu

725 730 735

<---

Claims (94)

1. A nucleic acid encoding a functional acidic α-glucosidase (GAA), wherein the nucleic acid has more than 90% sequence identity to a sequence selected from the group consisting of any of the sequences set forth as SEQ ID NOs: 1-5. 2. A nucleic acid encoding a functional GAA, wherein said nucleic acid has greater than 87% sequence identity to a sequence selected from the group consisting of any of the sequences shown as SEQ ID NO: 1-5, and contains a polyadenylation sequence that has reduced CpG content compared to the wild-type polyadenylation sequence located in the 3' direction of the specified nucleic acid. 3. A nucleic acid encoding a functional GAA according to claim 1 or 2, wherein said nucleic acid contains 20-10, 10-5 or 5-1 CpG dinucleotides or 0 CpG dinucleotides. 4. A nucleic acid encoding a functional GAA, wherein said nucleic acid has greater than 92% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-24. 5. A nucleic acid encoding a functional GAA, wherein the nucleic acid has a sequence selected from the group consisting of SEQ ID NO: 1-24. 6. The nucleic acid of claim 2 or 3, wherein said nucleic acid has greater than 88% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-5. 7. The nucleic acid of claim 2 or 3, wherein said nucleic acid has greater than 89% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-5. 8. The nucleic acid of claim 2 or 3, wherein said nucleic acid has greater than 90% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 6-15. 9. The nucleic acid of claim 2 or 3, wherein said nucleic acid has greater than 91% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-5. 10. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains less than 127 CpG dinucleotides. 11. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains less than 126 CpG dinucleotides. 12. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 126-120 CpG dinucleotides. 13. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 120-110 CpG dinucleotides. 14. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 110-100 CpG dinucleotides. 15. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 100-90 CpG dinucleotides. 16. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 90-80 CpG dinucleotides. 17. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 80-70 CpG dinucleotides. 18. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 70-60 CpG dinucleotides. 19. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 60-50 CpG dinucleotides. 20. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 50-40 CpG dinucleotides. 21. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 40-30 CpG dinucleotides. 22. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains approximately 30-20 CpG dinucleotides. 23. Nucleic acid according to any one of claims 1, 2, 4 or 6-8, wherein said nucleic acid contains less than 20 CpG dinucleotides. 24. Nucleic acid according to any one of claims 1-4 or 6-8, wherein said nucleic acid contains approximately 20-10 CpG dinucleotides. 25. Nucleic acid according to any one of claims 1-4 or 6-8, wherein said nucleic acid contains less than 10 CpG dinucleotides. 26. Nucleic acid according to any one of claims 1-4 or 6-8, wherein said nucleic acid contains approximately 10-5 CpG dinucleotides. 27. Nucleic acid according to any one of claims 1-4 or 6-8, wherein said nucleic acid contains 5 CpG dinucleotides. 28. Nucleic acid according to any one of claims 1-4 or 6-8, wherein said nucleic acid contains 4 CpG dinucleotides. 29. Nucleic acid according to any one of claims 1-4 or 6-8, wherein said nucleic acid contains 3 CpG dinucleotides. 30. Nucleic acid according to any one of claims 1-4 or 6-8, where the specified nucleic acid contains a 2-dinucleotide CpG. 31. Nucleic acid according to any one of claims 1-4 or 6-8, where the specified nucleic acid contains 1 CpG dinucleotide. 32. Nucleic acid according to any one of claims 1-4 or 6-8, wherein said nucleic acid contains 0 CpG dinucleotides. 33. The nucleic acid of claim 4, wherein said nucleic acid has greater than 93% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-24. 34. The nucleic acid of claim 4, wherein said nucleic acid has greater than 94% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-24. 35. The nucleic acid of claim 4, wherein said nucleic acid has greater than 95% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-24. 36. The nucleic acid of claim 4, wherein said nucleic acid has greater than 96% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-24. 37. The nucleic acid of claim 4, wherein said nucleic acid has greater than 97% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-24. 38. The nucleic acid of claim 4, wherein said nucleic acid has greater than 98% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-24. 39. The nucleic acid of claim 4, wherein said nucleic acid has greater than 99% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-24. 40. The nucleic acid of claim 4, wherein said nucleic acid has greater than 99.5% sequence identity to a sequence selected from the group consisting of any of the sequences set forth in SEQ ID NOs: 1-24. 41. An expression cassette comprising a nucleic acid according to any one of claims 1 to 40, operably linked to an expression control element. 42. The expression cassette according to claim 41, wherein said expression control element is located in the 5' direction of said nucleic acid. 43. The expression cassette of claim 41, further comprising a polyadenylation sequence located in the 3' direction of said nucleic acid. 44. The expression cassette of claim 42 or 43, wherein said expression control element or polyadenylation sequence has a reduced CpG content compared to the wild-type expression control element or polyadenylation sequence. 45. The expression cassette according to any one of claims 41-44, wherein said expression control element comprises an ApoE/hAAT enhancer/promoter sequence. 46. The expression cassette according to any one of claims 43 to 45, wherein said polyadenylation sequence comprises a bovine growth hormone (bGH) polyadenylation sequence. 47. The expression cassette of claim 45 or 46, wherein said ApoE/hAAT enhancer/promoter sequence or bGH polyadenylation sequence has a reduced CpG content compared to the ApoE/hAAT enhancer/promoter sequence or wild-type bGH polyadenylation sequence. 48. The expression cassette of claim 47, wherein said wild-type bGH polyadenylation sequence comprises the sequence SEQ ID NO: 27. 49. The expression cassette of claim 47, wherein said wild-type ApoE/hAAT enhancer/promoter sequence comprises SEQ ID NO: 28 or 29. 50. The expression cassette of claim 47, wherein said CpG-reduced bGH polyadenylation sequence comprises the sequence SEQ ID NO: 26. 51. The expression cassette according to any one of claims 41 to 50, further comprising an intron located between the 3' end of said expression control element and the 5' end of said nucleic acid. 52. The nucleic acid or expression cassette according to any one of claims 1 to 51, wherein said GAA contains the sequence shown as SEQ ID NO: 25. 53. An adeno-associated virus (AAV) vector for GAA expression, comprising the nucleic acid or expression cassette of claim 52. 54. The AAV vector according to claim 53, wherein said AAV vector contains: a) one or more AAV capsids and b) one or more AAV inverted terminal repeats (ITRs), wherein said AAV ITRs flank the 5' or 3' end of said nucleic acid or said expression cassette. 55. The AAV vector of claim 54, further comprising an intron located in the 5' or 3' direction of said one or more ITRs. 56. The AAV vector of claim 54 or 55, wherein at least one or more of said ITRs or said introns are modified to reduce CpG content. 57. The AAV vector according to any one of claims 53 to 56, wherein said AAV vector has a capsid serotype comprising a modified AAV VP1, VP2 and/or VP3 capsid or a variant AAV VP1, VP2 and/or VP3 capsid having 90% or more identity sequences in relation to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, Rh10, Rh74, AAV3B, AAV-2i8 or VP1, VP2 and/or VP3 sequences SEQ ID NO: 30, 31 or 32, or a capsid having 95% or more sequence identity to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, Rh10, Rh74, AAV3B, AAV-2i8, or VP1, VP2 and/or VP3 sequences SEQ ID NO: 30, 31 or 32, or a capsid having 100% sequence identity to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, Rh10, Rh74, AAV3B, AAV-2i8 or VP1, VP2 and/or VP3 sequences SEQ ID NO: 30, 31 or 32. 58. The AAV vector according to any one of claims 54-57, wherein said ITRs contain one or more ITRs from any of: AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, Rh10, Rh74 or AAV3B, AAV serotypes, or a combination thereof. 59. A pharmaceutical composition for treating a human in need of GAA, comprising a plurality of AAV vectors according to any one of claims 53 to 58 in a biocompatible carrier or excipient. 60. The pharmaceutical composition according to claim 59, further comprising empty AAV capsids. 61. The pharmaceutical composition of claim 60, wherein the ratio of said empty AAV capsids to said AAV vector is in the range of or about 100:1-50:1, about 50:1-25:1, about 25:1-10:1, approximately 10:1-1:1, approximately 1:1-1:10, approximately 1:10-1:25, approximately 1:25-1:50, or approximately 1:50-1:100. 62. The pharmaceutical composition of claim 60, wherein the ratio of said empty AAV capsids to said AAV vector is approximately 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9 :1 or 10:1. 63. Pharmaceutical composition according to any one of claims 60-62, additionally containing a surfactant. 64. A method of treating a person in need of acid α-glucosidase (GAA), comprising: (a) obtaining a nucleic acid or expression cassette according to any one of claims 1 to 52, or an AAV vector according to any of claims 53 to 58, or a pharmaceutical composition according to any of claims 59 to 63; And (b) administering an amount of said nucleic acid, expression cassette, AAV vector or pharmaceutical composition to said person, wherein said GAA is expressed in said person. 65. The method of claim 64, wherein said person has Pompe disease. 66. The method according to claim 64, wherein said person has an infantile form of Pompe disease. 67. The method of claim 64, wherein said person has late-onset Pompe disease. 68. The method of claim 64, wherein said person has glycogen storage disease (GSD). 69. The method according to claim 68, where the specified GSD is selected from: GSD type I (Gierke's disease), GSD type III (Forbes disease), GSD type IV (Andersen disease, amylopectinosis), GSD type V (McArdle disease), GSD type VI (Hers disease), GSD type VII (Tarui disease) and lethal congenital GSD of the heart. 70. The method according to any one of claims 64-69, wherein said AAV vector is administered to said individual intravenously, intraarterially, intracavitarily, intramucosally or via a catheter. 71. The method of any one of claims 64 to 70, wherein said GAA is expressed at elevated levels, optionally greater than 1% of GAA levels found in a GAA-neutral human. 72. The method of any one of claims 64 to 71, wherein said AAV vector is administered in the range of from about 1x108 to about 1x1014 vector genomes per kilogram (KG/kg) of weight of said individual. 73. The method according to any one of claims 64-72, wherein the specified method reduces, reduces or inhibits one or more symptoms of the specified need for GAA or the specified disease; or prevent or reduce the progression or worsening of one or more symptoms of a specified GAA need or a specified disease; or stabilize one or more symptoms of a specified GAA need or a specified disease; or improve one or more symptoms of a specified GAA need or a specified disease. 74. The method according to claim 73, where the specified one or more symptoms are selected from the group consisting of: problems with eating; no weight gain; poor head control; poor neck control; breathing problems; pulmonary infections; enlarged heart; thickening of the heart; heart defects; tongue enlargement; problems with swallowing; liver enlargement; low muscle strength; low muscle tone; weakness in the legs; weakness in the waist area; weakness in the hands; shortness of breath; problems with exercise; difficulty breathing during sleep; spinal curvature and ankylosis. 75. A cell containing a nucleic acid or expression cassette according to any one of claims 1 to 52, wherein the nucleic acid or expression cassette encodes a functional GAA. 76. An AAV vector-producing cell transfected with a nucleic acid or expression cassette according to any one of claims 53-58. 77. A method for producing an AAV vector according to any one of claims 53-58, including a. inserting an AAV vector genome containing said nucleic acid or expression cassette according to any one of claims 1 to 52 into a packaging helper cell; And b. culturing said helper cell under conditions to obtain said AAV vector. 78. A method for producing an AAV vector according to any one of claims 53-58, including a. incorporating said nucleic acid or expression cassette according to claims 1 to 52 into a packaging helper cell; And b. culturing said helper cells under conditions to obtain said AAV vector. 79. A cell or method according to any one of claims 75 to 78, wherein said cell includes mammalian cells. 80. A cell or method according to any one of claims 75-78, wherein said cell performs helper functions for packaging said vector into a viral particle. 81. A cell or method according to any one of claims 75-78, wherein said cell performs AAV helper functions. 82. The cell or method according to any one of claims 75-78, wherein said cell provides Rep and/or Cap proteins to AAV. 83. The cell or method according to any one of claims 75 to 78, wherein said cell is stably or transiently transfected using polynucleotides encoding Rep and/or Cap protein sequences. 84. The cell or method according to any one of claims 75-78, wherein said cell provides Rep78 and/or Rep68 proteins. 85. The cell or method of any one of claims 75 to 78, wherein said cell is stably or transiently transfected using sequences encoding Rep78 and/or Rep68 proteins. 86. A cell or method according to any one of claims 75 to 78, wherein said cell includes HEK-293 cells.

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