[go: up one dir, main page]

WO1987006261A1 - Systeme de conditionnement d'arn recombinant - Google Patents

Systeme de conditionnement d'arn recombinant Download PDF

Info

Publication number
WO1987006261A1
WO1987006261A1 PCT/GB1987/000249 GB8700249W WO8706261A1 WO 1987006261 A1 WO1987006261 A1 WO 1987006261A1 GB 8700249 W GB8700249 W GB 8700249W WO 8706261 A1 WO8706261 A1 WO 8706261A1
Authority
WO
WIPO (PCT)
Prior art keywords
rna
chimaeric
sequence
origin
virus
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB1987/000249
Other languages
English (en)
Inventor
Thomas Michael Aubrey Wilson
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Diatech Ltd
Original Assignee
Diatech Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Diatech Ltd filed Critical Diatech Ltd
Publication of WO1987006261A1 publication Critical patent/WO1987006261A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
    • C12N15/8206—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by physical or chemical, i.e. non-biological, means, e.g. electroporation, PEG mediated
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/00023—Virus like particles [VLP]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/00041—Use of virus, viral particle or viral elements as a vector
    • C12N2770/00043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

Definitions

  • the present invention relates to a recombinant - RNA packaging system based on helical, rod-shaped viruses.
  • Helical, rod-shaped ribonucleocapsids in particular plant viruses, consist of a hollow cylinder of viral coat protein encapsidating an RNA molecule.
  • the assembly of certain plant viruses has been studied quite extensively and it is known that in vitro TMV can be assembled from its constituent RNA molecule and coat protein in a spontaneous reaction. This reaction involves specific initiation at a nucleation site on the TMV RNA which is referred to as the origin of assembly sequence. Local sequence variations in the RNA, other than in the origin of assembly sequence, do not substantially affect the efficiency of assembly of the virus. It may be assumed that assembly of other helical, rod-shaped viruses is initiated and proceeds in a similar way.
  • RNA which includes the origin of assembly sequence of a helical, rod-shaped virus together with RNA coding for a foreign protein (referred to herein as a chimaeric RNA) , and that such a chimaeric RNA can be encapsidated by a preparation of the virus coat protein to form a virus-like particle
  • pseudovirus particles which include packaged RNA in a ribonuclease resistant form provide general vehicles for the protection, delivery and expression of said RNA.
  • the present invention provides a chimaeric RNA comprising the origin of assembly sequence of a helical, rod-shaped plant virus together with at least one sequence coding for a foreign protein.
  • the invention also provides a process for preparing a chimaeric RNA comprising the origin of assembly sequence of a helical, rod-shaped plant virus together with at least one sequence coding for a foreign protein, which comprises producing cloned cDNA copies of the RNA origin of assembly sequence and at least one cloned DNA sequence coding for a foreign protein, ligating the cloned DNA sequences in the correct orientation and transcribing the recombinant DNA in a suitable transcription vector system to produce the chimaeric RNA.
  • the invention also provides a pseudovirus particle comprising a chimaeric RNA which comprises the origin of assembly sequence of a helical rod-shaped plant virus together with at least one sequence coding for a foreign protein, encapsidated by the coat protein of the virus whose origin of assembly sequence is included in the chimaeric RNA.
  • the invention further provides a process for the production of a pseudovirus particle which comprises assembly of a chimaeric RNA comprising the origin of assembly sequence of a helical, rod-shaped plant virus together with at least one sequence coding for a foreign protein in a preparation of the coat protein of the virus whose origin of assembly sequence is included in the chimaeric RNA.
  • the invention provides a method for the expression of a heterologous protein in a host which comprises pseudo-infecting the said host with a pseudovirus particle comprising a chimaeric RNA which comprises the origin of assembly sequence of a helical rod-shaped plant virus together with a sequence coding for the foreign protein encapsidated by a coat protein of the virus whose origin of assembly sequence is included in the chimaeric RNA.
  • the pseudovirus particle according to the invention represents a vector which can be directly synthesised in large amounts and which contains RNA in a packaged form. Accordingly the pseudovirus particles can be used as a general means for the handling and storage of otherwise labile in vitro RNA transcripts.
  • the pseudovirus particles can be introduced into and expressed in a wide variety of hosts which extends far beyond the normal host for the virus on which the pseudovirus particle is based.
  • the pseudovirus particle represents an expression vector which., because of its packaging, is stable in the extra-cellular state and which is capable of "pseudo-infection" of plant cells which have not been subjected to any special treatment, i.e. the intact plant, as opposed to protoplasts, callus or suspension cultured cells.
  • pseudovirus particles derived from TMV has been observed in a plant species which is classified as a poor "subliminal" host for TMV. Accordingly the pseudovirus particles according to the invention based on plant viruses can be used as transient expression vectors in a wide range of hosts beyond the normal host of the plant virus concerned.
  • animal cells enopus laevis oocytes
  • animal cells are capable of uncoating pseudovirus particles according to the invention and expressing the encapsidated mRNA. Uncoating and expression is also possible in a cell free system derived from Escherichia coli cells. Accordingly the range of suitable hosts for pseudovirus particles according to the invention based on helical rod-shaped plant viruses appears very wide indeed and is not confined to plant cells
  • the invention can be applied to any helical, rod-shaped plant virus which assembles under the control of an origin of assembly sequence in a manner which is independent of the length and sequence of the remainder of the RNA.
  • TMV has been extensively characterised, including the origin of assembly sequence.
  • the invention can also be applied to other helical, rod-shaped plant viruses.
  • Suitable plant viruses apart from TMV include potato virus X which has the advantage that useful amounts of free coat protein for in vitro assembly of recombinant RNA transcripts are available in a workable form.
  • potato virus X has the advantage that useful amounts of free coat protein for in vitro assembly of recombinant RNA transcripts are available in a workable form.
  • the major mono-directional assembly mechanism is 5' — 3' and would require the potato virus X origin of assembly sequence upstream of the RNA sequence coding for the foreign protein of interest.
  • the term "origin of assembly sequence" of a helical rod-shaped plant virus means that part of the RNA sequence of the virus which is essential for a chimaeric RNA to be assembled into pseudovirus particles in the presence of the appropriate coat protein.
  • a 126 nucleotide sequence located between residues 5420 and 5546 from the 5 1 end of the TMV RNA molecule has been identified as the core of the sequence required for the nucleation of the TMV assembly with the residues 5313 to 5546 (the so-called extended region) also being implicated (see Goelet et al, Proc. Nat. Acad. Sci. U.S.A. 7£, 5818-5822 (1982) and Zimmern et al. Cell, JL1 455- 462 (1977)), although not all of this sequence is essential to effect assembly (Turner & Butler, Nuc. Acids Res, T4 9229 (1986) ) .
  • RNA origin of assembly sequence and of the cloned DNA sequences coding for a foreign protein, ligate the cloned DNA copies and transcribe the recombinant DNA in a suitable transcription system.
  • sequence coding for the foreign protein is also used in the form of DNA and both DNAs are inserted in a suitable orientation into a transcription vector.
  • Suitable vectors include the SP6 RNA polymerase plasmids pSP64 and pSP65 which are commercially available (Promega Biotec, Madison, WI, USA) and which contain a strong promoter for bacteriophage SP6 RNA polymerase.
  • Suitable plasmids include the dual promoter plasmids (e.g. pGEM 1-4 also available from Promega Biotec) which use SP6, T3 and/or T7 RNA polymerases.
  • pGEM 1-4 also available from Promega Biotec
  • any central foreign insert can be run off and packaged 3 ! — 5' in either the positive or negative (anti)- sense.
  • Addition of rho-independent transcription termination signals would probably enhance the overall yield of transcripts if arranged outside the motif described above since the template would no longer need to be linearized.
  • Transcripts can be 5'-capped, by using, for example, m 7 Gppp... as a primer for transcription, to prolong and enhance their cellular life and activity.
  • RNA transcripts In vitro packaging of the chimaeric RNA transcripts can be carried out using a prefabricated "disk" preparation of TMV coat protein under the assembly conditions published by Butler, J. Gen. Virol. 65. 253-279 (1984) and Durham, J. Mol. Biol. 67 289-305 (1972). Assembly can be monitored turbidometrically at 310 n using unlabelled transcripts, by recovery of icrococcal nuclease-resistant 32 P-labelled transcripts or by electron microscopy of negatively-stained nucleoprotein helices.
  • the original pseudovirus particles described above are essentially single round expression vector systems in the sense that they are not designed to replicate in "pseudo- infected" plants.
  • a pseudovirus particle based on TMV as described above should thus be capable of expression in any plant cell and as noted above the host range extends beyond plant cells. It may be possible to prepare pseudovirus particles according to the invention which are capable of replication but these will probably be more host-specific.
  • the essential elements of the vectors according to the invention can be used in conjunction with any other putative "vector/delivery" system to provide protective packaging for RNA constructs which are larger than otherwise tolerable in, for example, an isometric (spherical) nucleocapsid.
  • Figure 1 represents pSP64-derived constructs capable of directing the synthesis of specific RNA's containing the TMV origin of assembly sequence.
  • Figure 2 illustrates electrophoresis of linearized pSP64- derived RNA constructs before and after encapsidation, or following exposure to ribonuclease.
  • Figure 3 shows the results of electron microscopy of packaged in vitro transcripts.
  • Figure 4 shows the results of sucrose-density gradient fractionation of packaged, labelled SP6-transcripts.
  • Recombinant plasmids designated pSP64TMV, pSP64CT, pSP64LT, pSP64LRT, pJIIl and pJII2 which contain the TMV origin of assembly sequence (OAS) were constructed using the commercially available pSP64 plasmid (Promega Biotec) which contains the strong promoter for bacteriophage SP6 RNA polymerase. All plasmid constructs were grown in Escherichia coli strain DH1 and prepared using standard procedures as described by Maniatis, T. , Fritsch, E.F., and Sambrook. , J. in Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory (1982) .
  • the pSP64-derived plasmids are illustrated in Figure 1 in which the arrows represent the start of transcription by SP6 RNA polymerase.
  • a fragment corresponding to residues 5118-5550 of the total TMV (vulgare strain) RNA sequence and containing both the "core" (positions 5420-5546) and extended (position 5313- 5546) OAS region was supplied by Dr. P. Goelet as an Mspl fragment of TMV cDNA inserted into the AccI site of M13mp7 (Goelet and Karn, Gene 29., 331-342 (1984)).
  • the TMV OAS was excised as a BamHI fragment and subcloned into M13mpl0 (commercially available from New England Biolabs, see Messing, Methods in Enzymology, 101, 20 (1983)) from which a .clone containing the OAS, in the desired orientation for later 3' —).5' assembly, was selected by sequencing using the method of Sanger et al ., J. Mol. Biol. 143 161-178 (1980).
  • the TMV fragment was then excised from M13mpl0 by double-digestion with EcoRI and Sail and cloned into pSP64 to produce pSP64TMV. When linearized with EcoRI, this construct produces transcripts of 508 nucleotides (n) .
  • a cDNA sequence coding for calf preprochymosin was supplied as plasmid pSP64Chy82 + by Dr. A. Colman (University of Warwick) as a 1151 bp Bell fragment with Hindlll linkers, inserted into the Hindlll site of pSP64 (see Drummond et al, Nucleic Acid Res. 127375-7394 where the plasmid pSP64 Chy82 + is referred to on page 7377 as psp82 + ) .
  • This Hindlll fragment was transferred directly into the Hindlll site of pSP64TMV to produce pSP64CT.
  • the orientation was determined by restriction mapping. When linearized with Sad, this construct produces transcripts of 1659n, containing a 3'-OAS.
  • pUC8Lys + is a Hindlll-linkered fragment from K2 ys + (see Drummond et al. , Nucleic Acids Res. 13 7375-7394 (1985), which contains the chicken lysozyme gene which was inserted into pUC8 (commercially available from Bethesda Research Labs (see Vieira et al. , Gene JL9, 259 (1982)). This fragment was transferred to the Hindlll site of pSP64TMV to produce pSP64LT. When linearized with EcoRI, this construct produces transcripts of 993n and when linearized with BamHI it produces transcripts of 523n, lacking the TMV OAS.
  • Plasmid PSP64LRT identical to PSP64LT except for the addition of a 5.3kbp rDNA fragment between the lysozyme coding region and the TMV OAS.
  • the plasmid pJIIl is derived from pSP64TMV by linker conversion, wherein pSP64TMV is cut with Smal and ligated with
  • transcripts were incubated at 20°C with 40U/ml micrococcal nuclease and lmM C Cl2. Reactions were stopped by addition of EGTA to lOmM and SDS to 2% (w/v) prior to phenol/choloroform extraction and ethanol precipitation.
  • transcripts were analyzed by electrophoresis on a 1% (w/v) agarose/7.5% formaldehyde/MOPS denaturing gel system (Kreig et al.. , Nucleic Acid. Res. 12 - 7057-7070 (1984) ) :
  • tracks 1-4 represent the initial SP6-transcript (1) , the naked transcript digested with micrococcal nuclease (2) , the transcript incubated with TMV protein for 1 hr at 20°C and recovered without nuclease digestion (3) or following 30 min digestion with micrococcal nuclease (4) .
  • Tracks marked M represent SP6 transcripts of known size (235n, 683n, 1442n or 1784n) produced by linearizing pSP65 at known restriction sites for PvuII, Ddel, SinI or Seal respectively.
  • SP6 transcripts pSP64LT was linearized with EcoRI or BamHI. The latter enzyme removed a DNA fragment corresponding to the TMV OAS seguence (compare Fig. 2., tracks Bl & Cl) . 1.5 microgram of either template was incubated under standard (50 microlitre) reaction conditions (see Example 2) with 100 micro molar unlabelled rUTP and 10 micro Curies alpha-[ 32 p]-rUTP for 2hr initially with 15U SP6 RNA polymerase (Boehringer, Mannheim) . An additional 10U of polymerase were added after lhr.
  • Radiolabelled transcripts were recovered as described and two aliquots, each equivalent to 25% of the total yield of RNA, were packaged separately with TMV protein at an estimated protein:RNA ratio of 100:1.
  • One sample was stored on ice while the second was digested at 20°C with micrococcal nuclease (Boehringer, Mannheim) at 300U/ml in 3mM CaCl 2 . After 30min, EGTA was added to 5mM final concentration. All samples (in 100 microlitres) , including 12.5% aliquots of the original RNA transcripts, were loaded onto linear, 15-30% (w/v) DEP-treated sucrose density-gradients (5ml) , buffered with 0.1M Tris-HCl, pH8.0 at 5°C.
  • Sedimentation was from right to left in each case.
  • Fig.3D confirms the presence of adequate amounts of free TMV protein to complete the assembly
  • Fig.3G demonstrates the predominance of 70-90nm nucleoprotein rodlets.
  • Fig.2 track D4 (visible on original autoradiograph)
  • Fig.3C,F the majority appear to be only partially-coated to form 45-60nm rodlets
  • the efficiency of encapsidation of the chimaeric RNA transcripts can be estimated by (i) measuring the absolute concentration of rodlets observed in the electron microscope (Fig. 3) , (ii) calculation from the yield of radiolabelled transcripts recovered in nucleoprotein structures following micrococcal-nuclease digestion (Fig. 2, tracks A4-E4) , or (iii) sucrose density-gradient ultra-centrifugation of the assembled, radiolabelled transcripts, as shown in Fig. 4.
  • Fig. 3 The efficiency of encapsidation of the chimaeric RNA transcripts can be estimated by (i) measuring the absolute concentration of rodlets observed in the electron microscope (Fig. 3) , (ii) calculation from the yield of radiolabelled transcripts recovered in nucleoprotein structures following micrococcal-nuclease digestion (Fig. 2, tracks A4-E4) , or (iii) sucrose density-gradient ultra-centrifugation of the assembled, radiolabelled transcripts, as
  • RNA coding for a readily assayable protein CAT
  • buffer refers to 0.25M Tris-HCl, pH 7.4, containing lO M dithiothreitol and 2mM leupeptin.
  • Pseudovirus particles were prepared from Bglll linearized plasmid pJII2 by the method described in Example 2, except that the steps of 32 P-rUTP labelling and incubation of the naked or packaged transcripts at 20°C with micrococcal nuclease and CaCl 2 were omitted. Pseudovirus particles were also prepared from capped transcripts of linearized pJII2 using standard techniques. The majority of pseudovirus particles produced corresponded to the predicted length " for CAT pseudovirus particles of about 60nm.
  • Tobacco cells are natural hosts for TMV.
  • Tobacco mesophyll protoplasts were polyethylene glycol inoculated (Dawson et al. , Z. Naturforsch. C. Biosci. 33., 548 (1978)) with the following preparations 1) PEG alone 2) CAT mRNA
  • Samples 2 and 4 represent the mRNA transcripts from the line'arized plasmid pJII2 used to produce pseudovirus particles in accordance with Example 5 and samples 3 and 5 represent the pseudovirus particles themselves produced in accordance with Example 5.
  • Samples 2 - 5 received equivalent amounts of RNA on a weight basis. Following innoculation the protoplasts were incubated at 25°C for 20 hours. Protoplasts were removed from isotonic culture medium (Dawson et al. supra) by centrifuga ion, then resuspended and sonicated (10 sec) in an equal volume of buffer.
  • Pea (Pisu sativum L) is classified as one of the poorest "subliminal" hosts for TMV (Cheo, P.C. and Gerard, J.S. Phytopathology .61, 1010 (1971)).
  • CAT-expression in epidermal cells of Argenteum pea was investigated by inoculating pseudovirus particles or equivalent amounts of unencapsidated CAT mRNA constructs directly onto the leaf surface with silicon carbide [Carborundum 180 grit] as an abrasive.
  • the mutant Argenteum (Marx, J. Heredity 21413 (1982)) was used in view of its easily-peeled epidermis.
  • Example 5 The preparations tested were as for Example 5 except that buffer was used as a control. Other samples were applied so that equivalent amounts of RNA on a weight basis were used. Strips of epidermal cells were removed after 90 minutes and stored in liquid nitrogen (Shaw et al.. , Virology 148 326 (1986)) before being ground to a frozen powder. Lysed cells were resuspended in 300 microlitres of buffer. Cellular debris were removed and CAT assays performed as described in Example 5. 0.1 Unit of purified CAT was added to the sample inoculated with buffer as a reference.
  • Example 8 To determine whether pseudovirus particles could be uncoated and the mRNA expressed in animal cells Xenopus laevis oocytes were micro-injected separately with water as a control and with equivalent amounts of preparations 2 to 5 referred to in Example 5. Oocytes were also injected with the linearized plasmid DNA template containing the CAT coding sequence and the TMV origin of assembly to rule out any coupled transcription-translation activity. For further details of the methodology of oocyte injection see Colman in Transcription and Translation: A Practical Approach Ed. Hames et al IRL Press, Oxford (1984) pages 271-302.
  • the assay for CAT activity showed that the pseudovirus particles 3 and 5 produced CAT activity in the Xenopus oocyte system.
  • the Xenopus oocyte system responded more efficiently to non-encapsidated CAT mRNAs than to the corresponding pseudovirus particles.
  • a significant number of pseudovirus particles were disassembled and the resulting CAT mRNA expressed. This result suggests that the cytoplasm of animal cells includes suitable and sufficient machinery to disassemble the pseudovirus particles according to the invention.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Virology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Un ARN chimérique comprend l'origine de la séquence d'assemblage d'un virus en forme de tige hélicoïdale tel que le virus de la mosaïque du tabac avec au moins une séquence codant une protéine étrangère. L'ARN chimérique peut être obtenu en produisant des copies d'ADNc clonées de l'origine ARN de la séquence d'assemblage et des séquences d'ADN clonées codant une protéine étrangère, par ligation des séquences d'ADN clonées dans l'orientation correcte et transcription de l'ADN recombinant dans un système vectoriel de transcription appropriée. Lorsque l'ARN chimérique est assemblé dans une préparation de la protéine d'enrobage du virus dont l'origine de la séquence d'assemblage est incorporée dans l'ARN chimérique, une particule de pseudovirus est produite dans laquelle l'ARN chimérique est encapsulée par la protéine d'enrobage du virus. La particule de pseudovirus peut être utilisée comme moyen général pour protéger l'ARN ou comme vecteur d'expression de l'ARN dans une grande variété d'hôtes.
PCT/GB1987/000249 1986-04-11 1987-04-13 Systeme de conditionnement d'arn recombinant Ceased WO1987006261A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB868608850A GB8608850D0 (en) 1986-04-11 1986-04-11 Packaging system
GB8608850 1986-04-11

Publications (1)

Publication Number Publication Date
WO1987006261A1 true WO1987006261A1 (fr) 1987-10-22

Family

ID=10596044

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1987/000249 Ceased WO1987006261A1 (fr) 1986-04-11 1987-04-13 Systeme de conditionnement d'arn recombinant

Country Status (2)

Country Link
GB (1) GB8608850D0 (fr)
WO (1) WO1987006261A1 (fr)

Cited By (86)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994000588A1 (fr) * 1992-06-26 1994-01-06 British Technology Group Ltd. Systeme d'apport medicamenteux a base proteique
US5443969A (en) * 1992-10-29 1995-08-22 Rutgers University RNA packaging system
US5677124A (en) * 1996-07-03 1997-10-14 Ambion, Inc. Ribonuclease resistant viral RNA standards
US5766885A (en) * 1993-11-01 1998-06-16 Texas A & M University Potyvirus vectors for the expression of foreign genes
US5939262A (en) * 1996-07-03 1999-08-17 Ambion, Inc. Ribonuclease resistant RNA preparation and utilization
US6110466A (en) * 1991-04-19 2000-08-29 Axis Genetics Plc Modified plant viruses as vectors
US6232099B1 (en) 1994-10-18 2001-05-15 Scottish Crop Research Institute Method of producing a chimeric protein
US7148400B1 (en) * 1999-04-20 2006-12-12 Bayer Bioscience N.V. Methods and means for delivering inhibitory RNA to plants and applications thereof
WO2007020638A2 (fr) 2005-08-15 2007-02-22 Evogene Ltd. Procedes visant a augmenter la tolerance au stress abiotique et/ou la biomasse des plantes et plantes ainsi obtenues
WO2008010228A2 (fr) 2006-07-20 2008-01-24 Yeda Research And Development Co. Ltd. Organismes photosynthétiques, compositions et procédés de génération desdits organismes
US7385106B2 (en) 2000-01-24 2008-06-10 Ramot At Tel Aviv University Ltd. Plants tolerant of environmental stress conditions, methods of generating same and novel polynucleotide sequence utilized thereby
WO2008122980A2 (fr) 2007-04-09 2008-10-16 Evogene Ltd. Polynucléotides, polypeptides et procédés permettant d'augmenter la teneur en huile, la vitesse de croissance et la biomasse de plantes
WO2009083958A2 (fr) 2007-12-27 2009-07-09 Evogene Ltd. Peptides isolés, polynucléotides utiles pour modifier l'efficacité d'utilisation de l'eau, l'efficacité d'utilisation d'engrais, la tolérance aux stress biotiques/abiotiques, le rendement et la biomasse dans des plantes
WO2010049897A2 (fr) 2008-10-30 2010-05-06 Evogene Ltd. Polynucléotides et polypeptides isolés et procédés pour les utiliser pour augmenter le rendement, la biomasse, la vitesse de croissance, la vigueur, la teneur en huile, la tolérance au stress abiotique de plantes et l'efficacité d'utilisation de l'azote
WO2010076756A2 (fr) 2008-12-29 2010-07-08 Evogene Ltd. Polynucleotides, polypeptides codes par ceux-ci, et leurs procedes d'utilisation pour augmenter la tolerance au stress abiotique, la biomasse et/ou le rendement dans les plantes les exprimant
WO2010100595A2 (fr) 2009-03-02 2010-09-10 Evogene Ltd. Polynucléotides et polypeptides isolés, et procédés d'utilisation de ceux-ci pour augmenter un rendement végétal et/ou des caractéristiques agricoles
US7906705B2 (en) 2006-07-03 2011-03-15 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization, (A.R.O.), Volcani Center Polynucleotides and polypeptides encoded therefrom and methods of using same for increasing biomass in plants and plants generated thereby
WO2011048600A1 (fr) 2009-10-21 2011-04-28 Danziger Innovations Ltd. Génération de variations génotypiques dans des génomes de plantes par infection d'un gamète
EP2316950A1 (fr) 2000-03-27 2011-05-04 Technion Research and Development Foundation, Ltd. Complexes majeurs d'histocompatibilite monocatenaires de classe 1, constructions les codant et leurs methodes de production
WO2011067745A2 (fr) 2009-12-06 2011-06-09 Rosetta Green Ltd. Compositions et procédés d'amélioration de résistance de plantes au stress abiotique
EP2336330A2 (fr) 2004-06-14 2011-06-22 Evogene Ltd. Polynucléotides et polypeptides impliqués dans le développement de la fibre végétale et procédés permettant des les utiliser
WO2011080674A2 (fr) 2009-12-28 2011-07-07 Evogene Ltd. Polynucléotides isolés et polypeptides et procédés pour les utiliser afin d'augmenter le rendement des cultures, la biomasse, la vitesse de croissance, la vigueur, la teneur en huile, la tolérance au stress abiotique des plantes et l'efficacité d'utilisation de l'azote
US7977457B2 (en) 2006-05-19 2011-07-12 Teva Pharmaceutical Industries Ltd. Fusion proteins, uses thereof and processes for producing same
WO2011099006A2 (fr) 2010-02-11 2011-08-18 Yeda Research And Development Co. Ltd. Systèmes enzymatiques pour la fixation du carbone et procédés de production associés
EP2365087A2 (fr) 2003-05-22 2011-09-14 Evogene Ltd. Procédés d'augmentation de la tolérance du stress abiotique des plantes et plantes ainsi produites
EP2383345A1 (fr) 2006-12-20 2011-11-02 Evogene Ltd. Polynucléotides et polypeptides impliqués dans le développement de fibres végétales et procédés d'utilisation
WO2011135527A2 (fr) 2010-04-28 2011-11-03 Evogene Ltd. Polynucléotides et polypeptides isolés et procédés d'utilisation de ceux-ci pour l'augmentation du rendement végétal et/ou des caractéristiques agricoles
WO2011158242A2 (fr) 2010-06-16 2011-12-22 Futuragene Israel Ltd. Plantes résistantes aux organismes nuisibles
WO2012007919A2 (fr) 2010-07-15 2012-01-19 Technion Research & Development Foundation Ltd. Construction d'acide nucléique destinée à accroître la tolérance au stress abiotique dans des végétaux
WO2012007945A2 (fr) 2010-07-12 2012-01-19 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization, (A.R.O.), Volcani Center Polynucléotides isolés et procédés et plantes les utilisant pour la régulation de l'acidité des plantes
WO2012032520A1 (fr) 2010-09-07 2012-03-15 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Translecture d'acétylcholinestérase (ache-r) pour le traitement ou la prévention de la maladie de parkinson
EP2441840A1 (fr) 2005-07-18 2012-04-18 Protalix Ltd. Administration mucosale ou enterale de macromolecules biologiquement actives
WO2012059922A2 (fr) 2010-11-03 2012-05-10 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Plantes transgéniques à rendements de saccharification améliorés, et procédé pour les générer
WO2012098537A1 (fr) 2011-01-20 2012-07-26 Protalix Ltd. Construction d'acide nucléique pour l'expression d'une alpha-galactosidase chez des plantes et des cellules végétales
WO2012117406A2 (fr) 2011-03-02 2012-09-07 Futuragene Israel Ltd. Plantes transgéniques résistantes aux bactéries
WO2012156976A1 (fr) 2011-05-16 2012-11-22 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Procédés de préparation d'artémisinine dans des plantes non-hôtes et vecteurs destinés à une utilisation dans celles-ci
US8455717B2 (en) 2004-09-29 2013-06-04 Collplant Ltd. Collagen producing plants and methods of generating and using same
EP2599790A1 (fr) 2007-11-26 2013-06-05 Yissum Research Development Company of The Hebrew University of Jerusalem Compositions comprenant des polypeptides fibreux et des polysachharides
WO2013088438A1 (fr) 2011-12-11 2013-06-20 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization, (A.R.O.), Volcani Center Procédés de modulation de conductance stomatique et constructions d'expression de plante pour exécuter ces procédés
WO2013114373A1 (fr) 2012-02-01 2013-08-08 Protalix Ltd. Formulations liquides inhalables d'adnase i
WO2013121405A1 (fr) 2012-02-19 2013-08-22 Protalix Ltd. Formes galéniques unitaires administrées par voie orale et utilisations de celles-ci pour le traitement de la maladie de gaucher
WO2014033723A1 (fr) 2012-09-03 2014-03-06 A.B. Seeds Ltd. Procédé d'amélioration de la tolérance au stress abiotique des plantes et plantes ainsi produites
EP2716654A1 (fr) 2005-10-24 2014-04-09 Evogene Ltd. Polypeptides isolés, polynucléotides codant pour ces derniers, plantes transgéniques exprimant ces derniers et méthodes utilisant ces derniers
WO2014136114A1 (fr) 2013-03-06 2014-09-12 Protalix Ltd. Polypeptides inhibiteurs de tnf alpha, polynucléotides codant pour ceux-ci, cellules exprimant ceux-ci et leurs procédés de production
WO2014136117A1 (fr) 2013-03-06 2014-09-12 Protalix Ltd. Utilisation de cellules végétales exprimant un inhibiteur du polypeptide tnf alpha en thérapie
EP2816117A2 (fr) 2004-09-29 2014-12-24 Collplant Ltd. Collagène recombinant produit dans une plante
WO2015118547A1 (fr) 2014-02-10 2015-08-13 Protalix Ltd. Méthode de maintien de la stabilité d'une maladie chez un sujet atteint de la maladie de gaucher
WO2015118183A1 (fr) * 2014-02-10 2015-08-13 Amptec Gmbh Arn enveloppé
EP2910638A2 (fr) 2007-07-24 2015-08-26 Evogene Ltd. Polynucléotides, polypeptides codés correspondants et procédés d'utilisation associés permettant d'améliorer la tolérance au stress abiotique et/ou la biomasse et/ou le rendement dans des plantes les exprimant
EP2936976A1 (fr) 2008-04-21 2015-10-28 Danziger Innovations Ltd. Vecteurs d'expression de virus de plante et leur utilisation pour générer des variations génotypiques dans des génomes de plantes
WO2015189693A1 (fr) 2014-06-12 2015-12-17 King Abdullah University Of Science And Technology Édition ciblée de génome de plante à médiation virale à l'aide du système crispr/cas9
WO2016038617A1 (fr) 2014-09-11 2016-03-17 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Procédés de production de mogrosides et compositions les comprenant et leurs utilisations
EP3020410A1 (fr) 2008-04-18 2016-05-18 Collplant Ltd. Procédés de génération et d'utilisation de procollagène
WO2016079739A2 (fr) 2014-11-20 2016-05-26 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Compositions et procédés de production de polypeptides présentant un schéma de glycosylation modifié dans des cellules végétales
WO2016084084A1 (fr) 2014-11-27 2016-06-02 Danziger Innovations Ltd. Construction d'acide nucléique pour l'édition de génome
EP3072972A2 (fr) 2008-08-18 2016-09-28 Evogene Ltd. Polypeptides et polynucléotides isolés utiles pour augmenter l'efficacité de l'utilisation de l'azote, la tolérance au stress abiotique, le rendement et la biomasse de plantes
WO2017125931A1 (fr) 2016-01-21 2017-07-27 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Plantes parthénocarpiques et procédés de production de celles-ci
EP3238534A2 (fr) 2009-06-10 2017-11-01 Evogene Ltd. Polynucléotides et polypeptides isolés et leurs procédés d'utilisation pour augmenter l'efficacité d'utilisation, le rendement, la vitesse de croissance, la vigueur, la biomasse, la teneur en huile et/ou la tolérance au stress abiotique de l'azote
WO2018220579A1 (fr) 2017-05-31 2018-12-06 Tropic Biosciences UK Limited Compositions et procédés permettant d'augmenter l'extractibilité de solides en provenance de grains de café
WO2018220581A1 (fr) 2017-05-31 2018-12-06 Tropic Biosciences UK Limited Compositions et procédés pour l'augmentation de la durée de conservation de la banane
WO2018220582A1 (fr) 2017-05-31 2018-12-06 Tropic Biosciences UK Limited Méthodes de sélection de cellules comprenant des événements d'édition de génome
WO2018224861A1 (fr) 2017-06-07 2018-12-13 International Rice Research Institute Augmentation de la production de semences hybrides par un taux de croisement plus élevé chez des plantes graminacées stériles cytoplasmiques et matériaux et procédés associés
US10184131B2 (en) 2012-02-06 2019-01-22 A.B. Seeds Ltd. Isolated polynucleotides expressing or modulating microRNAs or targets of same, transgenic plants comprising same and uses thereof
WO2019106641A2 (fr) 2017-12-03 2019-06-06 Seedx Technologies Inc. Systèmes et procédés de tri de graines
WO2019106638A1 (fr) 2017-12-03 2019-06-06 Seedx Technologies Inc. Systèmes et procédés de tri de graines
WO2019106639A1 (fr) 2017-12-03 2019-06-06 Seedx Technologies Inc. Systèmes et procédés de tri de graines
WO2019211854A1 (fr) 2018-05-03 2019-11-07 Collplant Holdings Ltd. Charges dermiques et applications de celles-ci
WO2019211750A1 (fr) 2018-05-01 2019-11-07 Tropic Biosciences UK Limited Compositions et procédés de réduction de la teneur en caféine dans les grains de café
WO2019234750A1 (fr) 2018-06-07 2019-12-12 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Procédés de régénération et de transformation de cannabis
WO2019234754A1 (fr) 2018-06-07 2019-12-12 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Constructions d'acide nucléique et procédés d'utilisation de celles-ci
WO2020008412A1 (fr) 2018-07-04 2020-01-09 Ukko Inc. Procédés de ré-épitopage de protéines de blé et leur utilisation pour le traitement de maladie cœliaque
WO2020183419A1 (fr) 2019-03-14 2020-09-17 Tropic Biosciences UK Limited Introduction d'une activité de silençage sur des molécules d'arn dysfonctionnelles et modification de leur spécificité vis-à-vis d'un gène d'intérêt
WO2020183416A1 (fr) 2019-03-14 2020-09-17 Tropic Biosciences UK Limited Production d'arndb dans des cellules végétales pour la protection antiparasitaire via le silencage génique
WO2020183414A2 (fr) 2019-03-14 2020-09-17 Tropic Biosciences UK Limited Modification de la spécificité de molécules d'arn non codantes pour le silençage de l'expression génique dans des cellules eucaryotes
WO2021001784A1 (fr) 2019-07-04 2021-01-07 Ukko Inc. Alpha gliadine sans épitope et son utilisation pour la gestion de la maladie coeliaque et de la sensibilité au gluten
WO2021019536A1 (fr) 2019-07-30 2021-02-04 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Méthodes de régulation de la synthèse de cannabinoïdes chez la plante ou dans des cellules et plantes et cellules ainsi produites
WO2021100034A1 (fr) 2019-11-19 2021-05-27 Protalix Ltd. Élimination de constructions à partir de cellules transformées
US11104948B2 (en) 2014-02-10 2021-08-31 Vela Operations Singapore Pte. Ltd. NGS systems control and methods involving the same
WO2021202513A1 (fr) 2020-03-31 2021-10-07 Elo Life Systems Modulation des gènes de la voie endogène des mogrosides chez la pastèque et d'autres cucurbitacées
WO2022038536A1 (fr) 2020-08-18 2022-02-24 International Rice Research Institute Procédés d'augmentation des taux de croisement de gramineae
WO2022074646A1 (fr) 2020-10-05 2022-04-14 Protalix Ltd. Cellules végétales inactivées de type dicer
WO2022087527A1 (fr) 2020-10-23 2022-04-28 Elo Life Systems, Inc. Procédés de production de vanilliers à arôme et production agronomique améliorées
WO2022115498A1 (fr) 2020-11-26 2022-06-02 Ukko Inc. Sous-unité de gluténine modifiée à poids moléculaire élevé et ses utilisations
WO2022226316A1 (fr) 2021-04-22 2022-10-27 Precision Biosciences, Inc. Compositions et procédés pour générer des plantes mâles stériles
US11555199B2 (en) 2017-09-19 2023-01-17 Tropic Biosciences UK Limited Modifying the specificity of plant non-coding RNA molecules for silencing gene expression
WO2023036984A1 (fr) 2021-09-13 2023-03-16 Plantibodies Organisme génétiquement modifié pour la production de protéines recombinées

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0067553A2 (fr) * 1981-05-27 1982-12-22 National Research Council Of Canada Vecteur à base d'ARN de virus de plante ou une partie de celui-ci, procédé pour sa production, et une méthode de production d'un produit dérivé de gène, à l'aide de celui-ci
EP0153154A1 (fr) * 1984-02-14 1985-08-28 Lubrizol Genetics Inc. Vecteur de transfert
EP0194809B1 (fr) * 1985-03-07 1991-03-13 Lubrizol Genetics Inc. Vecteur de transformation d'ARN
JPH0641486A (ja) * 1991-03-19 1994-02-15 Arakawa Chem Ind Co Ltd 印刷インキ用バインダーおよびその製造法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0067553A2 (fr) * 1981-05-27 1982-12-22 National Research Council Of Canada Vecteur à base d'ARN de virus de plante ou une partie de celui-ci, procédé pour sa production, et une méthode de production d'un produit dérivé de gène, à l'aide de celui-ci
EP0153154A1 (fr) * 1984-02-14 1985-08-28 Lubrizol Genetics Inc. Vecteur de transfert
EP0194809B1 (fr) * 1985-03-07 1991-03-13 Lubrizol Genetics Inc. Vecteur de transformation d'ARN
JPH0641486A (ja) * 1991-03-19 1994-02-15 Arakawa Chem Ind Co Ltd 印刷インキ用バインダーおよびその製造法

Non-Patent Citations (11)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, Volume 102, 1985, (Columbus, Ohio, US), D'ANN ROCHON et al.: "TMV Coat Protein Encapsidates Specific Species of Host RNA Both in Vivo and in Vitro", see page 341, Abstract 163542s, UCLA Symp. Mol. Cell. Biol., New Ser. 1985, 22 (Cell. Mol. Biol. Plant Stress), 435-46 *
CHEMICAL ABSTRACTS, Volume 103, 1985, (Columbus, Ohio, US), T. WILSON et al.: "Nucleocapsid Disassembly and Early Gene Expression by Positive-Strand RNA Viruses", see page 307, Abstract 19483b, J. Gen. Virol. 1985, 66 (6), 1201-7 *
CHEMICAL ABSTRACTS, Volume 103, No. 9, September 1985, (Columbus, Ohio, US), see page 181, Abstract 66069f, & JP, A, 6041486 (Kirin Brewery Co., LTD) K March 1985 *
CHEMICAL ABSTRACTS, Volume 104, 1986 (Columbus, Ohio, US), G.P. LOMONOSSOFF et al.: "Structure and in Vitro Assembly of Tobacco Mosaic Virus", see page 299, Abstract 31414h, Mol. Plant Virol. 1985, 1, 43-83 *
CHEMICAL ABSTRACTS, Volume 104, 1986, (Columbus, Ohio, US), M. TABLER et al.: "Infectivity Studies on Different Potato Spindle Tuber Viroid (PSTV) RNAs Synthesized in Vitro with the SP6 Transcription System", see pages 137-138, Abstract 29605c, EMBO J. 1985, 4(9), 2191-9 *
CHEMICAL ABSTRACTS, Volume 105, 1986, (Columbus, Ohio, US), D.L. NUSS: "Engineering a Plant RNA Virus for Expression of Foreign Genetic Sequences", see page 165, Abstract 147138w, BioEssays 1986, 4(3), 133-4 *
CHEMICAL ABSTRACTS, Volume 106, 1987, (Columbus, Ohio, US), D.E. SLEAT et al.: "Packaging of Recombinant RNA Molecules into Pseudovirus Particles Directed by the Origin-of-Assembly Sequence from Tobacco Mosaic Virus RNA", see page 158, Abstract 62188h, Virology 1986, 155(2), 299-308 *
CHEMICAL ABSTRACTS, Volume 80, 1974, (Columbus, Ohio, US), D.R. BLACK et al.: "Structure and Infectivity of Picornaviral RNA Encapsidated by Cowpea Chlorotic Mottle Virus Protein", see page 130, Abstract 35047z, J. Virol. 1973, 12(6), 1209-15 *
CHEMICAL ABSTRACTS, Volume 87, 1977, (Columbus, Ohio, US), D. ZIMMERN et al.: "The Isolation of Tobacco Mosaic Virus RNA Fragments Containing the Origin for Viral Assembly", see pages 246-247, Abstract 129216a, Cell (Cambridge, Mass.) 1977, 11 (3), 455-62 cited in the application *
Nature, Volume 219, 17 August 1968, S. ROGERS et al.: "Use of Viruses as Carriers of Added Genetic Information", pages 749-751 see the whole document *
Proc. Natl. Acad. Sci. USA, Volume 83, March 1986, W.O. DAWSON et al.: "cDNA Cloning of the Complete Genome of Tobacco Mosaic Virus and Production of Infectious Transcripts", pages 1832-1836 see the Abstract *

Cited By (122)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7208655B1 (en) 1991-04-19 2007-04-24 The Dow Chemical Company Modified plant viruses as vectors
US6110466A (en) * 1991-04-19 2000-08-29 Axis Genetics Plc Modified plant viruses as vectors
US6884623B1 (en) 1991-04-19 2005-04-26 The Dow Chemical Company Modified plant viruses as vectors of heterologous peptides
WO1994000588A1 (fr) * 1992-06-26 1994-01-06 British Technology Group Ltd. Systeme d'apport medicamenteux a base proteique
KR100240515B1 (ko) * 1992-06-26 2000-01-15 말콤 카터, 리차드 케이쓰 퍼시 단백질을 기초로 한 전달 시스템
GB2268492B (en) * 1992-06-26 1996-04-17 British Tech Group Protein based delivery system
US6159728A (en) * 1992-06-26 2000-12-12 Btg International Limited RNA bacteriophage-based delivery system
US5443969A (en) * 1992-10-29 1995-08-22 Rutgers University RNA packaging system
EP0683821A4 (fr) * 1992-10-29 1996-03-27 Univ Rutgers Systeme de compactage d'arn.
US5766885A (en) * 1993-11-01 1998-06-16 Texas A & M University Potyvirus vectors for the expression of foreign genes
US6232099B1 (en) 1994-10-18 2001-05-15 Scottish Crop Research Institute Method of producing a chimeric protein
US5939262A (en) * 1996-07-03 1999-08-17 Ambion, Inc. Ribonuclease resistant RNA preparation and utilization
US6214982B1 (en) 1996-07-03 2001-04-10 Ambion Inc Ribonuclease resistant RNA preparation and utilization
US6399307B1 (en) 1996-07-03 2002-06-04 Ambion, Inc. Methods of quantifying viral load in an animal with a ribonuclease resistant RNA preparation
US5677124A (en) * 1996-07-03 1997-10-14 Ambion, Inc. Ribonuclease resistant viral RNA standards
US7033749B2 (en) 1996-07-03 2006-04-25 Ambion, Inc. Ribonuclease resistant RNA preparation and utilization
US5919625A (en) * 1996-07-03 1999-07-06 Ambion, Inc. Ribonuclease resistant viral RNA standards
US7148400B1 (en) * 1999-04-20 2006-12-12 Bayer Bioscience N.V. Methods and means for delivering inhibitory RNA to plants and applications thereof
US7385106B2 (en) 2000-01-24 2008-06-10 Ramot At Tel Aviv University Ltd. Plants tolerant of environmental stress conditions, methods of generating same and novel polynucleotide sequence utilized thereby
EP2316950A1 (fr) 2000-03-27 2011-05-04 Technion Research and Development Foundation, Ltd. Complexes majeurs d'histocompatibilite monocatenaires de classe 1, constructions les codant et leurs methodes de production
EP2365087A2 (fr) 2003-05-22 2011-09-14 Evogene Ltd. Procédés d'augmentation de la tolérance du stress abiotique des plantes et plantes ainsi produites
EP2336330A2 (fr) 2004-06-14 2011-06-22 Evogene Ltd. Polynucléotides et polypeptides impliqués dans le développement de la fibre végétale et procédés permettant des les utiliser
EP2343373A1 (fr) 2004-06-14 2011-07-13 Evogene Ltd. Polynucléotides et polypeptides impliqués dans le développement de la fibre végétale et procédés permettant des les utiliser
US8455717B2 (en) 2004-09-29 2013-06-04 Collplant Ltd. Collagen producing plants and methods of generating and using same
EP2816117A2 (fr) 2004-09-29 2014-12-24 Collplant Ltd. Collagène recombinant produit dans une plante
US10626408B2 (en) 2004-09-29 2020-04-21 Collplant Ltd. Collagen producing plants and methods of generating and using same
US9783816B2 (en) 2004-09-29 2017-10-10 Collplant Ltd. Collagen producing plants and methods of generating and using same
EP3088528A1 (fr) 2004-09-29 2016-11-02 Collplant Ltd. Plantes produisant du collagene et procedes de generation et d'utilisation de celles-ci
EP2441840A1 (fr) 2005-07-18 2012-04-18 Protalix Ltd. Administration mucosale ou enterale de macromolecules biologiquement actives
EP2484768A2 (fr) 2005-07-18 2012-08-08 Protalix Ltd. Administration mucosale ou enterale de macromolecules biologiquement actives
WO2007020638A2 (fr) 2005-08-15 2007-02-22 Evogene Ltd. Procedes visant a augmenter la tolerance au stress abiotique et/ou la biomasse des plantes et plantes ainsi obtenues
EP2995194A1 (fr) 2005-08-15 2016-03-16 Evogene Ltd. Procédés permettant d'augmenter la tolérance au stress abiotique et/ou la biomasse dans des plantes et plantes ainsi produites
EP2716654A1 (fr) 2005-10-24 2014-04-09 Evogene Ltd. Polypeptides isolés, polynucléotides codant pour ces derniers, plantes transgéniques exprimant ces derniers et méthodes utilisant ces derniers
US7977457B2 (en) 2006-05-19 2011-07-12 Teva Pharmaceutical Industries Ltd. Fusion proteins, uses thereof and processes for producing same
US7906705B2 (en) 2006-07-03 2011-03-15 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization, (A.R.O.), Volcani Center Polynucleotides and polypeptides encoded therefrom and methods of using same for increasing biomass in plants and plants generated thereby
WO2008010228A2 (fr) 2006-07-20 2008-01-24 Yeda Research And Development Co. Ltd. Organismes photosynthétiques, compositions et procédés de génération desdits organismes
EP2383345A1 (fr) 2006-12-20 2011-11-02 Evogene Ltd. Polynucléotides et polypeptides impliqués dans le développement de fibres végétales et procédés d'utilisation
WO2008122980A2 (fr) 2007-04-09 2008-10-16 Evogene Ltd. Polynucléotides, polypeptides et procédés permettant d'augmenter la teneur en huile, la vitesse de croissance et la biomasse de plantes
EP2910638A2 (fr) 2007-07-24 2015-08-26 Evogene Ltd. Polynucléotides, polypeptides codés correspondants et procédés d'utilisation associés permettant d'améliorer la tolérance au stress abiotique et/ou la biomasse et/ou le rendement dans des plantes les exprimant
EP2599790A1 (fr) 2007-11-26 2013-06-05 Yissum Research Development Company of The Hebrew University of Jerusalem Compositions comprenant des polypeptides fibreux et des polysachharides
WO2009083958A2 (fr) 2007-12-27 2009-07-09 Evogene Ltd. Peptides isolés, polynucléotides utiles pour modifier l'efficacité d'utilisation de l'eau, l'efficacité d'utilisation d'engrais, la tolérance aux stress biotiques/abiotiques, le rendement et la biomasse dans des plantes
EP3020410A1 (fr) 2008-04-18 2016-05-18 Collplant Ltd. Procédés de génération et d'utilisation de procollagène
EP2936976A1 (fr) 2008-04-21 2015-10-28 Danziger Innovations Ltd. Vecteurs d'expression de virus de plante et leur utilisation pour générer des variations génotypiques dans des génomes de plantes
EP3616504A2 (fr) 2008-08-18 2020-03-04 Evogene Ltd. Polypeptides et polynucléotides isolés utiles pour augmenter l'efficacité de l'utilisation de l'azote, la tolérance au stress abiotique, le rendement et la biomasse de plantes
EP3072972A2 (fr) 2008-08-18 2016-09-28 Evogene Ltd. Polypeptides et polynucléotides isolés utiles pour augmenter l'efficacité de l'utilisation de l'azote, la tolérance au stress abiotique, le rendement et la biomasse de plantes
WO2010049897A2 (fr) 2008-10-30 2010-05-06 Evogene Ltd. Polynucléotides et polypeptides isolés et procédés pour les utiliser pour augmenter le rendement, la biomasse, la vitesse de croissance, la vigueur, la teneur en huile, la tolérance au stress abiotique de plantes et l'efficacité d'utilisation de l'azote
WO2010076756A2 (fr) 2008-12-29 2010-07-08 Evogene Ltd. Polynucleotides, polypeptides codes par ceux-ci, et leurs procedes d'utilisation pour augmenter la tolerance au stress abiotique, la biomasse et/ou le rendement dans les plantes les exprimant
EP3000889A2 (fr) 2009-03-02 2016-03-30 Evogene Ltd. Polynucléotides et polypeptides isolés, et procédés d'utilisation de ceux-ci pour augmenter un rendement végétal et/ou des caractéristiques agricoles
WO2010100595A2 (fr) 2009-03-02 2010-09-10 Evogene Ltd. Polynucléotides et polypeptides isolés, et procédés d'utilisation de ceux-ci pour augmenter un rendement végétal et/ou des caractéristiques agricoles
EP3626051A2 (fr) 2009-06-10 2020-03-25 Evogene Ltd. Polynucléotides et polypeptides isolés et leurs procédés d'utilisation pour augmenter l'efficacité d'utilisation, le rendement, la vitesse de croissance, la vigueur, la biomasse, la teneur en huile et/ou la tolérance au stress abiotique de l'azote
EP3238534A2 (fr) 2009-06-10 2017-11-01 Evogene Ltd. Polynucléotides et polypeptides isolés et leurs procédés d'utilisation pour augmenter l'efficacité d'utilisation, le rendement, la vitesse de croissance, la vigueur, la biomasse, la teneur en huile et/ou la tolérance au stress abiotique de l'azote
WO2011048600A1 (fr) 2009-10-21 2011-04-28 Danziger Innovations Ltd. Génération de variations génotypiques dans des génomes de plantes par infection d'un gamète
US9562235B2 (en) 2009-12-06 2017-02-07 A.B. Seeds Ltd. MicroRNA compositions and methods for enhancing plant resistance to abiotic stress
WO2011067745A2 (fr) 2009-12-06 2011-06-09 Rosetta Green Ltd. Compositions et procédés d'amélioration de résistance de plantes au stress abiotique
WO2011080674A2 (fr) 2009-12-28 2011-07-07 Evogene Ltd. Polynucléotides isolés et polypeptides et procédés pour les utiliser afin d'augmenter le rendement des cultures, la biomasse, la vitesse de croissance, la vigueur, la teneur en huile, la tolérance au stress abiotique des plantes et l'efficacité d'utilisation de l'azote
EP3056569A2 (fr) 2009-12-28 2016-08-17 Evogene Ltd. Polynucléotides et polypeptides isolés et procédés d'utilisation de ceux-ci pour augmenter le rendement, la biomasse, la vitesse de croissance, la vigueur, la teneur en huile, la tolérance au stress abiotique et l'efficacité d'utilisation de l'azote de plantes
WO2011099006A2 (fr) 2010-02-11 2011-08-18 Yeda Research And Development Co. Ltd. Systèmes enzymatiques pour la fixation du carbone et procédés de production associés
WO2011135527A2 (fr) 2010-04-28 2011-11-03 Evogene Ltd. Polynucléotides et polypeptides isolés et procédés d'utilisation de ceux-ci pour l'augmentation du rendement végétal et/ou des caractéristiques agricoles
WO2011158242A2 (fr) 2010-06-16 2011-12-22 Futuragene Israel Ltd. Plantes résistantes aux organismes nuisibles
WO2012007945A2 (fr) 2010-07-12 2012-01-19 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization, (A.R.O.), Volcani Center Polynucléotides isolés et procédés et plantes les utilisant pour la régulation de l'acidité des plantes
WO2012007919A2 (fr) 2010-07-15 2012-01-19 Technion Research & Development Foundation Ltd. Construction d'acide nucléique destinée à accroître la tolérance au stress abiotique dans des végétaux
WO2012032520A1 (fr) 2010-09-07 2012-03-15 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Translecture d'acétylcholinestérase (ache-r) pour le traitement ou la prévention de la maladie de parkinson
WO2012059922A2 (fr) 2010-11-03 2012-05-10 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Plantes transgéniques à rendements de saccharification améliorés, et procédé pour les générer
WO2012098537A1 (fr) 2011-01-20 2012-07-26 Protalix Ltd. Construction d'acide nucléique pour l'expression d'une alpha-galactosidase chez des plantes et des cellules végétales
WO2012117406A2 (fr) 2011-03-02 2012-09-07 Futuragene Israel Ltd. Plantes transgéniques résistantes aux bactéries
WO2012156976A1 (fr) 2011-05-16 2012-11-22 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Procédés de préparation d'artémisinine dans des plantes non-hôtes et vecteurs destinés à une utilisation dans celles-ci
WO2013088438A1 (fr) 2011-12-11 2013-06-20 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization, (A.R.O.), Volcani Center Procédés de modulation de conductance stomatique et constructions d'expression de plante pour exécuter ces procédés
WO2013114374A1 (fr) 2012-02-01 2013-08-08 Protalix Ltd. Polypeptides d'adnase i, polynucléotides codant ces derniers, procédés de production d'adnase i et utilisations de celle-ci en thérapie
WO2013114371A1 (fr) 2012-02-01 2013-08-08 Protalix Ltd. Formulation en poudre sèche de dnase i
US9603907B2 (en) 2012-02-01 2017-03-28 Protalix Ltd. Dry powder formulations of dNase I
US9603906B2 (en) 2012-02-01 2017-03-28 Protalix Ltd. Inhalable liquid formulations of DNase I
WO2013114373A1 (fr) 2012-02-01 2013-08-08 Protalix Ltd. Formulations liquides inhalables d'adnase i
US10184131B2 (en) 2012-02-06 2019-01-22 A.B. Seeds Ltd. Isolated polynucleotides expressing or modulating microRNAs or targets of same, transgenic plants comprising same and uses thereof
WO2013121405A1 (fr) 2012-02-19 2013-08-22 Protalix Ltd. Formes galéniques unitaires administrées par voie orale et utilisations de celles-ci pour le traitement de la maladie de gaucher
WO2014033723A1 (fr) 2012-09-03 2014-03-06 A.B. Seeds Ltd. Procédé d'amélioration de la tolérance au stress abiotique des plantes et plantes ainsi produites
WO2014136113A1 (fr) 2013-03-06 2014-09-12 Protalix Ltd. Polypeptides chimériques, polynucléotides codant pour ceux-ci, cellules exprimant ceux-ci et leurs procédés de production
WO2014136117A1 (fr) 2013-03-06 2014-09-12 Protalix Ltd. Utilisation de cellules végétales exprimant un inhibiteur du polypeptide tnf alpha en thérapie
WO2014136114A1 (fr) 2013-03-06 2014-09-12 Protalix Ltd. Polypeptides inhibiteurs de tnf alpha, polynucléotides codant pour ceux-ci, cellules exprimant ceux-ci et leurs procédés de production
WO2015118183A1 (fr) * 2014-02-10 2015-08-13 Amptec Gmbh Arn enveloppé
JP2017505143A (ja) * 2014-02-10 2017-02-16 アンプテック ゲゼルシャフト ミット ベシュレンクテル ハフツング インコーテッドｒｎａ
US11104948B2 (en) 2014-02-10 2021-08-31 Vela Operations Singapore Pte. Ltd. NGS systems control and methods involving the same
WO2015118547A1 (fr) 2014-02-10 2015-08-13 Protalix Ltd. Méthode de maintien de la stabilité d'une maladie chez un sujet atteint de la maladie de gaucher
WO2015189693A1 (fr) 2014-06-12 2015-12-17 King Abdullah University Of Science And Technology Édition ciblée de génome de plante à médiation virale à l'aide du système crispr/cas9
US11584936B2 (en) 2014-06-12 2023-02-21 King Abdullah University Of Science And Technology Targeted viral-mediated plant genome editing using CRISPR /Cas9
WO2016038617A1 (fr) 2014-09-11 2016-03-17 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Procédés de production de mogrosides et compositions les comprenant et leurs utilisations
US11697819B2 (en) 2014-11-20 2023-07-11 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd Compositions and methods for producing polypeptides with a modified glycosylation pattern in plant cells
WO2016079739A2 (fr) 2014-11-20 2016-05-26 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Compositions et procédés de production de polypeptides présentant un schéma de glycosylation modifié dans des cellules végétales
WO2016084084A1 (fr) 2014-11-27 2016-06-02 Danziger Innovations Ltd. Construction d'acide nucléique pour l'édition de génome
WO2017125931A1 (fr) 2016-01-21 2017-07-27 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Plantes parthénocarpiques et procédés de production de celles-ci
WO2018220582A1 (fr) 2017-05-31 2018-12-06 Tropic Biosciences UK Limited Méthodes de sélection de cellules comprenant des événements d'édition de génome
WO2018220581A1 (fr) 2017-05-31 2018-12-06 Tropic Biosciences UK Limited Compositions et procédés pour l'augmentation de la durée de conservation de la banane
WO2018220579A1 (fr) 2017-05-31 2018-12-06 Tropic Biosciences UK Limited Compositions et procédés permettant d'augmenter l'extractibilité de solides en provenance de grains de café
EP4461813A2 (fr) 2017-05-31 2024-11-13 Tropic Biosciences UK Limited Compositions et procédés pour augmenter la durée de conservation de banane
WO2018224861A1 (fr) 2017-06-07 2018-12-13 International Rice Research Institute Augmentation de la production de semences hybrides par un taux de croisement plus élevé chez des plantes graminacées stériles cytoplasmiques et matériaux et procédés associés
US12275939B2 (en) 2017-09-19 2025-04-15 Tropic Biosciences UK Limited Modifying the specificity of plant non-coding RNA molecules for silencing gene expression
EP4170029A2 (fr) 2017-09-19 2023-04-26 Tropic Biosciences UK Limited Modification de la spécificité de molécules d'arn non codantes végétales pour le silençage de l'expression génique
US12331295B2 (en) 2017-09-19 2025-06-17 Tropic Biosciences UK Limited Modifying the specificity of plant non-coding RNA molecules for silencing gene expression
US11555199B2 (en) 2017-09-19 2023-01-17 Tropic Biosciences UK Limited Modifying the specificity of plant non-coding RNA molecules for silencing gene expression
WO2019106639A1 (fr) 2017-12-03 2019-06-06 Seedx Technologies Inc. Systèmes et procédés de tri de graines
WO2019106641A2 (fr) 2017-12-03 2019-06-06 Seedx Technologies Inc. Systèmes et procédés de tri de graines
WO2019106638A1 (fr) 2017-12-03 2019-06-06 Seedx Technologies Inc. Systèmes et procédés de tri de graines
WO2019211750A1 (fr) 2018-05-01 2019-11-07 Tropic Biosciences UK Limited Compositions et procédés de réduction de la teneur en caféine dans les grains de café
WO2019211854A1 (fr) 2018-05-03 2019-11-07 Collplant Holdings Ltd. Charges dermiques et applications de celles-ci
EP4446363A2 (fr) 2018-05-03 2024-10-16 CollPlant Ltd. Charges dermiques et leurs applications
US12186449B2 (en) 2018-05-03 2025-01-07 Collplant Ltd. Dermal fillers and applications thereof
US11801329B2 (en) 2018-05-03 2023-10-31 Collplant Ltd. Dermal fillers and applications thereof
WO2019234750A1 (fr) 2018-06-07 2019-12-12 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Procédés de régénération et de transformation de cannabis
WO2019234754A1 (fr) 2018-06-07 2019-12-12 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Constructions d'acide nucléique et procédés d'utilisation de celles-ci
WO2020008412A1 (fr) 2018-07-04 2020-01-09 Ukko Inc. Procédés de ré-épitopage de protéines de blé et leur utilisation pour le traitement de maladie cœliaque
WO2020183419A1 (fr) 2019-03-14 2020-09-17 Tropic Biosciences UK Limited Introduction d'une activité de silençage sur des molécules d'arn dysfonctionnelles et modification de leur spécificité vis-à-vis d'un gène d'intérêt
WO2020183414A2 (fr) 2019-03-14 2020-09-17 Tropic Biosciences UK Limited Modification de la spécificité de molécules d'arn non codantes pour le silençage de l'expression génique dans des cellules eucaryotes
WO2020183416A1 (fr) 2019-03-14 2020-09-17 Tropic Biosciences UK Limited Production d'arndb dans des cellules végétales pour la protection antiparasitaire via le silencage génique
WO2021001784A1 (fr) 2019-07-04 2021-01-07 Ukko Inc. Alpha gliadine sans épitope et son utilisation pour la gestion de la maladie coeliaque et de la sensibilité au gluten
WO2021019536A1 (fr) 2019-07-30 2021-02-04 The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Center) Méthodes de régulation de la synthèse de cannabinoïdes chez la plante ou dans des cellules et plantes et cellules ainsi produites
WO2021100034A1 (fr) 2019-11-19 2021-05-27 Protalix Ltd. Élimination de constructions à partir de cellules transformées
WO2021202513A1 (fr) 2020-03-31 2021-10-07 Elo Life Systems Modulation des gènes de la voie endogène des mogrosides chez la pastèque et d'autres cucurbitacées
WO2022038536A1 (fr) 2020-08-18 2022-02-24 International Rice Research Institute Procédés d'augmentation des taux de croisement de gramineae
WO2022074646A1 (fr) 2020-10-05 2022-04-14 Protalix Ltd. Cellules végétales inactivées de type dicer
WO2022087527A1 (fr) 2020-10-23 2022-04-28 Elo Life Systems, Inc. Procédés de production de vanilliers à arôme et production agronomique améliorées
WO2022115498A1 (fr) 2020-11-26 2022-06-02 Ukko Inc. Sous-unité de gluténine modifiée à poids moléculaire élevé et ses utilisations
WO2022226316A1 (fr) 2021-04-22 2022-10-27 Precision Biosciences, Inc. Compositions et procédés pour générer des plantes mâles stériles
WO2023036984A1 (fr) 2021-09-13 2023-03-16 Plantibodies Organisme génétiquement modifié pour la production de protéines recombinées

Also Published As

Publication number Publication date
GB8608850D0 (en) 1986-05-14

Similar Documents

Publication Publication Date Title
WO1987006261A1 (fr) Systeme de conditionnement d'arn recombinant
Meshi et al. In vitro transcription of infectious RNAs from full-length cDNAs of tobacco mosaic virus
Krappa et al. Identification of the very early transcribed baculovirus gene PE-38
Danthinne et al. The 3'untranslated region of satellite tobacco necrosis virus RNA stimulates translation in vitro
US6051753A (en) Figwort mosaic virus promoter and uses
Sleat et al. Packaging of recombinant RNA molecules into pseudovirus particles directed by the origin-of-assembly sequence from tobacco mosaic virus RNA
US5443969A (en) RNA packaging system
KR19990067271A (ko) 재조합체 센다이 바이러스
JPH08505289A (ja) 遺伝子導入植における組換メッセンジャーｒｎａのウィルス増幅関連出願のクロスリファレンス
AU2002250064B2 (en) A bi-directional dual promoter complex with enhanced promoter activity for transgene expression in eukaryotes
EP0781347A1 (fr) Produit de recombinaison par fusion de genes de peptides lytiques d'ubiquitine, produits en derivant et leur procede d'obtention
Osteryoung et al. Poly (A) tail length of a heat shock protein RNA is increased by severe heat stress, but intron splicing is unaffected
JPH0231683A (ja) Ｔ‐ｄｎａ７８０遺伝子の転写活性化要素
HUP0001817A2 (hu) Chlorella víruspromóterek
Copertino et al. A group III twintron encoding a maturase-like gene excises through lariat intermediates
KR101596229B1 (ko) 식물에 있어서 이종유래 단백질의 발현 향상을 위하여 5'(5'-ｕｔｒ) 내에 최적화된 리더 작용기를 가진 인공 ｄｎａ 서열
Chiang et al. Infectivity assays of in vitro and in vivo transcripts of papaya ringspot potyvirus
Van der Vossen et al. Role of the 5′ leader sequence of alfalfa mosaic virus RNA 3 in replication and translation of the viral RNA
JPS619288A (ja) ペプチド類の製法
Rosenberg et al. T7 RNA polymerase can direct expression of influenza virus cap-binding protein (PB2) in Escherichia coli
CN109879946B (zh) 白木香AsWRKY44转录因子及其应用
JP3055787B2 (ja) キュウリモザイクウイルスの外皮蛋白遺伝子
EP1181372B1 (fr) Vecteurs viraux d'expression pour les plantes
CN112592923A (zh) Ires序列、ires序列的应用和多顺反子表达载体
US6869780B2 (en) Nodavirus-like DNA expression vector and uses therefor

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): GB JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE FR GB IT LU NL SE