WO1995025806A2 - Packaging-deficient lentiviruses - Google Patents
Packaging-deficient lentiviruses Download PDFInfo
- Publication number
- WO1995025806A2 WO1995025806A2 PCT/GB1995/000663 GB9500663W WO9525806A2 WO 1995025806 A2 WO1995025806 A2 WO 1995025806A2 GB 9500663 W GB9500663 W GB 9500663W WO 9525806 A2 WO9525806 A2 WO 9525806A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- vector
- packaging
- virus
- vector according
- sequence
- Prior art date
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
- 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
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
-
- 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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/15011—Lentivirus, not HIV, e.g. FIV, SIV
- C12N2740/15041—Use of virus, viral particle or viral elements as a vector
- C12N2740/15043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- 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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/15011—Lentivirus, not HIV, e.g. FIV, SIV
- C12N2740/15051—Methods of production or purification of viral material
- C12N2740/15052—Methods of production or purification of viral material relating to complementing cells and packaging systems for producing virus or viral particles
Definitions
- This invention relates to lentiviruses, recombinant DNA technology and novel vectors for gene transfer. Background to the Invention
- lentiviruses a subgroup of retroviruses
- the genomic RNA species is the full-length unspliced message which is also capable of translation, encoding the gag and pol genes of the virus.
- Lentiviruses are able to specifically encapsidate their genomic RNA, because unique cis-acting signals that are present within this RNA enable differentiation from other viral RNA species.
- a number of RNA regions in retroviruses are reviewed by Schlesinger et al (1994), Seminars in Virology ,5:39-49, and have been identified as containing packaging signals that enable encapsidation (packaging) of genomic RNA into a virion particle.
- the cis-acting messages involved are usually in the form of sequence and structural motifs and are found in the 5' coding regions close to the splice donor (from deletion mutagenesis studies) .
- RNA protein interactions involved in packaging are poorly understood.
- the evidence suggests that an interaction of a RNA motif or motifs, with a component of the gag or gag /pol polyproteins is likely.
- GCG three base motif identified by Konings et al (1992) J. Virol j66:632-640
- This motif is often found in regions of lentiviral RNA (e.g. MPMV, SMRV-H) that are predicted to be unfolded. Therefore, whilst this motif may have an accessory function, the evidence so far suggests that it is not a major packaging signal.
- the packaging regions of various retroviruses have undergone further secondary structure RNA modelling using biochemical analysis and free energy minimisation studies in conjunction with phylogenetic studies. Harrison and Lever (1992) J.
- Virol 16:4144-4153 report a study involving the HIV-1 lentivirus, showing RNA encapsidation to be far more complex in this particular retroviru ⁇ subgroup than in other retroviruses such as avian and murine viruses. Apparently, packaging signals both within and outside of the 5' leader sequence are required for encapsidation.
- a packaging-defective lentivirus can be generated, by inactivating the known packaging signals for use as a vaccine. This has been achieved in HIV-1 by Richardson et al (1993) J. Virol. 67:3997-4005; see also WO-A-9317118 and the commonly-owned copending US Patent Application Serial No. 08/295,737, filed August 26, 1994, the contents of which are incorporated herein by reference.
- the non- infectious virion protein coat (containing antigenic epitopes) would then be presented to the host immune system to confer protective immunity without risk of infection, as the lentivirus lacks the viral RNA that gives rise to pathogenicity. It is envisaged that 'subunit' type vaccines could be designed on the basis of further study of the antigenic epitopes to which the immune response is directed.
- Lentiviral infections persist in cells of the macrophage/monocyte series and, in this immunologically privileged site, they are difficult to eradicate. Furthermore, pharmacological approaches to ameliorating lentiviral infection have proved difficult, due to the close relationship between virus and host processes. Even inhibitors of the virus-specific enzyme reverse transcriptase have demonstrable effects on host cell polymerases. Furthermore, lentiviruses can achieve a latent state in host cells, requiring lifelong therapeutic treatment which causes significant problems of drug toxicity due to accumulation.
- heterologous RNA in which a lentiviral packaging signal is incorporated, could be packaged into its corresponding virion particle (instead of the normal viral RNA) .
- This heterologous RNA may encode a gene which could then be specifically targeted to cells, using the hybrid virion, which are susceptible to a specific lentiviral infection.
- Carrier particles and their use for gene therapy are the subject of, inter alia, WO-A-9204916 and WO-A-9211846. Carriers and methods described there may be suitable for the purposes of the present invention. Summary of the Invention
- the present invention is based at least in part on the investigation of a stable secondary structure model for the 5' leader region of the prototype D-type retrovirus Mason Pfizer Monkey Virus, that is presented 3' of the primer- binding site.
- a stem-loop structure in the region has been identified, which from other studies has been shown to be important for genomic RNA encapsidation.
- the structure involves a highly stable stem of 5 G:C pairs terminating in a heptaloop.
- Comparison with a predicted structure for Squirrel Monkey Retrovirus with MPMV shows a structure with an identical stem and a common ACC motif in the loop. Free energy studies of secondary structure in nine other retroviruses predict stem loops which have similar GAYC motifs.
- the present invention utilises the consensus sequence GAYC as a unique packaging signal when enclosed in a stem-loop with a positioning 5' to the gag initiation codon and where this motif is not involved in another known interaction, for example, the complementary sequence of the tRNA primer binding site in the leader sequence. If that motif is already base-pairing with another RNA such as the tRNA primer, then it is unlikely to be involved in packaging; however, if it is uninvolved in any known interaction and is in the appropriate position, it appears that the motif is involved. According to the present invention, therefore, a vector is capable of producing a lentivirus protein, but not of packaging lentivirus RNA.
- oligonucleotides including the GAYC motif, or a complementary sequence are used to compete with viral proteins that interact with the RNA.
- the competition serves to prevent packaging of the viral RNA.
- a novel recombinant molecule for use as a vector may be generated.
- This may comprise nucleotides corresponding to the packaging nucleotides of a lentivirus, a heterologous gene and, flanking the packaging nucleotides and the heterologous gene, sequences including the .GAYC motif sufficient for packaging, reverse transcription and integration of the vector into target cells, to enable expression of the heterologous gene.
- This aspect of the invention is based on the realisation that lentiviruses are the only retroviruses able to integrate their DNA into non-dividing cells, increasing the potential target population for gene transfer. The property seems to reside in the matrix protein which probably has a nuclear localisation signal and may transport ONA through the nuclear membrane.
- incorporasation of this protein into particles will enhance nuclear transport of introduced genetic sequences. Integration may require an integrase enzyme. Alternatively, if the DNA includes suitable homologous sequences it may recombine with chromosomal DNA by homologous recombination. This process would both excise any unwanted gene and insert the correct one.
- the targeted gene could encode anti-sense expressing gene specifically targeted at an MW or HIV-2 sequence or at a cellular sequence involved in MW or HIV-2 replication.
- the lentiviral inhibitory gene or genes (multiple antiviral genes could be contained in one or multiple vectors delivered to a cell) might be expressed constitutively in the cell.
- the gene might be engineered such that is under the control of MW or HIV-2 regulatory genes, e.g. tat gene, whereby it would remain transcriptionally silent until activated by an incoming lentivirus.
- a recombinant DNA molecule comprises a gene which is functionally dependent on a metal, e.g. under the control of a non-physiological metal-dependent promoter. This is based in part on the realisation that the control of expression of genes may be affected by upstream promoter sequences. Some of these are metal-dependent, e.g. the MT2 promoter; transcription can then be activated by the appropriate cation. Further, a metal may cause activation of an enzyme that controls expression.
- Lentiviruses to which this invention relate include MPMV, and others. Description of the Invention
- RNA secondary structure To probe for RNA secondary structure, in vitro transcribed RNAs were either digested with structure- specific enzymes or were modified with structure-specific biochemicals.
- the transcribed RNAs were generated from the 5' leader region of MPMV excised from the proviral clone pSHRMl ⁇ [Rhee et al (1990) Eur. J. Bioch. 7:305-318] using the SphI site at position 567 and the Sa ⁇ l site at 1561.
- the numbering is that of RESIVMPC (Genbank accession no. M12349) . This fragment was ligated into the same sites in the expression vector pGem-4Z (Promega, Southampton, UK) and RNA was transcribed from the T7 promoter.
- RNA samples were digested with DNAse 1, then were dissociated and reannealed by heating to 65 ⁇ C for 5 ins and cooling to 25 ⁇ C over 20 minutes.
- RNAs were then divided into 2 ⁇ g aliquots and were modified with one of the following agents: Cobra venom RNase VI, (Pharmacia), hydroxymethylpsoralen (psoralen) (Sigma) , 2-keto-3-ethoxybutyraldehyde (kethoxal) (United States Biochemical) , RNAse Tl (Boehringer Mannheim) , and dimethyl sulphate (DMS) (Fluka) .
- Cobra venom RNase VI (Pharmacia)
- psoralen hydroxymethylpsoralen
- kethoxal 2-keto-3-ethoxybutyraldehyde
- RNAse Tl Boehringer Mannheim
- DMS dimethyl sulphate
- Photoreactions with psoralen were carried out under a 366 nm germicidal ultraviolet lamp. Samples were irradiated for 30 mins at room temperature in Eppendorf tubes at a distance of 20-30 mm from a Sylvania G875 tube light.
- RNAs were extracted with phenol/chloroform and 10 ng of a synthetic oligonucleotide primer was added in avian myeloblastosis virus (AMV) reverse transcriptase (RT) buffer (Pro ega) .
- Oligonucleotide primers which were used were: 5 , -TTG/CCC/CAT/ATC/CGA/GCG/C-3 , from bases 898 to 880. 5'-CCC/CGT/GTC/TTT/AAA/GCC/-3' from bases 937 to 939 and 5'-TAT/GGT/TCC/CTC/TTG/CGG/-3' from bases 1042 to 1025.
- RNAs were dissociated by heating the solution to 70 ⁇ C for 5 minutes. The solutions were then immediately stored on ice. It was found that the primer annealed when the solutions were incubated with the extension mix. Extension analyses were carried out by adding 1 unit of AMV RT (Promega) , a final cone of 1 mM each dATP, dTTP and dGTP and 1 ⁇ l [ ⁇ 32 P]dATP. These extension mixtures were incubated at 42°C for 1 hour. cDNAs were precipitated under ethanol, and redissolved in 1 x TE buffer.
- RNA in solution is in a dynamic state, and that alternative structures of the same regions exist simultaneously in a population of homologous molecules. Any individual RNA molecule may alternate between different conformations. This leads to some ambiguity in interpretation of biochemical data, as single stranded modifiers will be able to attack regions of RNA even if they are only transiently unpaired. A very stable double helix will however be much less vulnerable to single-stranded modification than a region which has no nearby complementary sequence.
- Psoralen intercalates in base-paired regions, and forms covalent adducts with pyrimidines, notably uridines when irradiated with ultra-violet light at 366 nm [Youvan and Hearst (1982) Anal. Biochemistry 119:86-89 r and Bachellerie et al (1981) Nucleic Acids Res. 2:2207-2222].
- Psoralen is found to intercalate particularly at the end of helices or in mismatches within helices [Leffers et al (1988) J. Mol. Biol. 104. / 507-522].
- Kethoxal modifies unpaired G residues, and DMS modifies A's and C's at positions which interfere with reverse transcription.
- RNAse Tl cleaves single-stranded RNA at G residues.
- Fig. 1 gives a comparison of predicted stem loops in the 5' leader regions of 11 different retroviruses.
- the murine structures which are boxed together were proposed by Tounekti et al (1992), J. Mol. Biol. 223.:205-220.
- the MoMuLV structure was first proposed by Alford et al (1991) , Virology 183:611-619. Sequence variations for SRV-l and 2 are indicated on the MPMV stem loop. The position of the gag AUG relative to the last C of the ACC motif is indicated for each retrovirus. In the 847-853 loop of the MPMV structure, there is a triplet ACC (849 to 851) which is conserved between MPMV and SMRV-H.
- SRV-1 and SRV-2 the purine is conserved.
- Free energy predictions for B-type viruses and a series of distantly related retroviruses including gibbon-ape leukaemia virus (GALV) , [AKR murine leukaemia virus (AKV) , FBJ-murine osteosarcoma virus (FBJ-MSV) , Friend spleen focus-forming virus (SFFV) , FBR-MSV and caprine arthritis and encephalitis virus (CAEV) , show similar motifs in stem loop structures in analogous positions relative to the gag initiation codon ( Figure 3) which conform to the general pattern GAYC (motif) . This is also found in bovine leukaemia virus (BLV) .
- the MPMV motif RCC is also seen in SNV and feline leukaemia virus (FeLV) at position 784.
- the stem and loop 878-898 containing the gag initiation codon are conserved between MPMV, SRV-1 and SRV-2 and SMRV and can also adopt a similar structure.
- the evidence strongly suggests that the GAYC motif may represent a common structural and sequence packaging motif in these viruses. Deletion mutations involving the region between the major splice site (nucleotides 303 to 304) and the gag initation codon (nucleotide 489) of the Maedi-Visna virus (MW) , which afflicts sheep, were constructed - see Figure 2.
- Figure 3 shows deletions in an analogous region between nucleotides 493 and 534 in the untranslated leader sequence in Human Immunodeficiency Virus type 2 (HIV-2) . These resulted in a similar packaging defective phenotype.
- the given nucleotide numbering is as in Guyader et al (1987) Nature 326:662-669.
- a second packaging signal will be required in both of these viruses to enable encapsidation of heterologous RNA.
- This is located at the 3' end of the genome in a region adjacent to the ⁇ is-acting sequence through which the rev gene product acts.
- the rev responsive sequence will in part be different from the second packaging signal.
- the invention thus allows the construction of a replication-defective MW or HIV-2. It allows the production of viral particles devoid of viral nucleic acid. These could be used as vaccines. It allows the production of particles into which RNA containing the necessary packaging signals of MW or HIV-2 can be packaged. These latter RNAs can contain heterologous genes and thus MW and HIV-2 can be developed as gene vectors for delivery of genes to cells for which these viruses are tropic.
- the invention allows the use of the intrinsic property of these two viruses to integrate a DNA copy of the genetic material carried within the virus into a target cell even if that cell is not undergoing mitosis.
- a packaging-defective MW or HIV-2 leads to the production of MW or HIV-2 virus particles which are non-infectious but which have all the correct MW or HIV-2 proteins assembled in their native form. They are therefore suitable for use as a vaccine.
- a similar approach might subsequently be used for other lentiviral disease of domestic animals such as Equine Infectious Anaemia virus and Caprine Arthritis-Encephalitis virus.
- Vaccine formulations may be of conventional nature.
- the desired gene can be targeted to the appropriate cell, e.g. by using a particle or vector. A dose of the appropriate cation can be targeted to the same cell (particles) .
- the cation may be, for example, cadmium (which is rare, non- physiological, and toxic if in the wrong place at the wrong concentration) .
- the double targeting step provides a desirable degree of safety, cell-targeting specificity and also flexible control of transcription, by giving doses of cation-containing particle when desired.
- the foreign gene may be, for example, for a ribozyme.
- Ribozymes are RNA enzymes capable of cleaving RNA target sequences. They are potential antiviral agents. Their function may be to switch off expression of other genes, e.g. oncogenes. Ribozymes can be encoded on plasmids grown in bacteria. Bacterial growth may be made ribozyme-dependent, i.e.
- Ribozymes are divalent cation-dependent, probably for stability rather than at the active site. By including in the medium for growth high concentrations of a particular non-physiological cation, e.g. strontium, ribozymes may be evolved which work optimally or only in the presence of the cation. An antiviral/anti-oncogene ribozyme is then made, and into it is incorporated the cation-dependent sequence.
- the ribozyme may now be delivered to the target cell, followed by the strontium or other cation.
- the delivery of each may be achieved using particles, for example by a technique described in WO-A-9204916 or WO-A-9211846.
- the ribozyme cannot be activated in cells without the cation, and therefore will not inhibit physiological processes, e.g. a normal short burst of expression of the oncogene.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Public Health (AREA)
- Physics & Mathematics (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Biophysics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Plant Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Biochemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
Claims
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP95912349A EP0752000A1 (en) | 1994-03-24 | 1995-03-24 | Packaging-deficient lentiviruses |
JP7524505A JPH09510361A (en) | 1994-03-24 | 1995-03-24 | Packaging defective lentivirus |
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB9405875A GB9405875D0 (en) | 1994-03-24 | 1994-03-24 | Recombinant molecules and their use in gene transfer |
GB9405876A GB9405876D0 (en) | 1994-03-24 | 1994-03-24 | Modified virus and its use as a vector |
GBGB9425026.3A GB9425026D0 (en) | 1994-12-09 | 1994-12-09 | Packaging-defective vectors |
GB9405875.7 | 1994-12-09 | ||
GB9405876.5 | 1994-12-09 | ||
GB9425026.3 | 1994-12-09 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO1995025806A2 true WO1995025806A2 (en) | 1995-09-28 |
WO1995025806A3 WO1995025806A3 (en) | 1996-01-04 |
Family
ID=27267117
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/GB1995/000663 WO1995025806A2 (en) | 1994-03-24 | 1995-03-24 | Packaging-deficient lentiviruses |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP0752000A1 (en) |
JP (1) | JPH09510361A (en) |
WO (1) | WO1995025806A2 (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0871459A1 (en) * | 1995-10-06 | 1998-10-21 | The Salk Institute For Biological Studies | Vector and method of use for nucleic acid delivery to non-dividing cells |
WO1999015641A1 (en) * | 1997-09-24 | 1999-04-01 | The Regents Of The University Of California | Non-primate lentiviral vectors and packaging systems |
US6790657B1 (en) | 1999-01-07 | 2004-09-14 | The United States Of America As Represented By The Department Of Health And Human Services | Lentivirus vector system |
WO2013037841A3 (en) * | 2011-09-12 | 2013-05-23 | Centre National De La Recherche Scientifique (C.N.R.S) | Chimeric non-integrating lentiviral genomes as vaccines against hiv-1 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1993017118A2 (en) * | 1992-02-28 | 1993-09-02 | Syngenix Limited | Defective packaging non-oncoviral vectors based on mpmv and hiv |
WO1993020220A1 (en) * | 1992-03-27 | 1993-10-14 | Whitehead Institute For Biomedical Research | Non-infectious hiv particles and uses therefor |
WO1994003622A1 (en) * | 1992-07-31 | 1994-02-17 | Imperial College Of Science, Technology & Medicine | D-type retroviral vectors, based on mpmv |
-
1995
- 1995-03-24 EP EP95912349A patent/EP0752000A1/en not_active Withdrawn
- 1995-03-24 WO PCT/GB1995/000663 patent/WO1995025806A2/en not_active Application Discontinuation
- 1995-03-24 JP JP7524505A patent/JPH09510361A/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1993017118A2 (en) * | 1992-02-28 | 1993-09-02 | Syngenix Limited | Defective packaging non-oncoviral vectors based on mpmv and hiv |
WO1993020220A1 (en) * | 1992-03-27 | 1993-10-14 | Whitehead Institute For Biomedical Research | Non-infectious hiv particles and uses therefor |
WO1994003622A1 (en) * | 1992-07-31 | 1994-02-17 | Imperial College Of Science, Technology & Medicine | D-type retroviral vectors, based on mpmv |
Non-Patent Citations (3)
Title |
---|
EMBO J., vol.13, pages 713 - 726 S. YANG AND H.M. TEMIN; 'A double hairpin structure is necessary for the efficient encapsidation of spleen necrosis virus retroviral DNA' * |
J. VIROL., vol.66, pages 632 - 640 D.A.M. KONINGS ET AL.; 'Novel GACG- hairpin pair motif in the 5' untranslated region of type C retroviruses related to murine leukemia virus' cited in the application * |
VIROLOGY, vol.189, pages 786 - 791 R.G. VILE ET AL.; 'Identification of a generalised packaging sequence for D-type retroviruses and generation of a D-type retroviral vector' * |
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0871459A1 (en) * | 1995-10-06 | 1998-10-21 | The Salk Institute For Biological Studies | Vector and method of use for nucleic acid delivery to non-dividing cells |
EP0871459A4 (en) * | 1995-10-06 | 2001-09-05 | Salk Inst For Biological Studi | Vector and method of use for nucleic acid delivery to non-dividing cells |
WO1999015641A1 (en) * | 1997-09-24 | 1999-04-01 | The Regents Of The University Of California | Non-primate lentiviral vectors and packaging systems |
AU747438B2 (en) * | 1997-09-24 | 2002-05-16 | Regents Of The University Of California, The | Non-primate lentiviral vectors and packaging systems |
US6555107B2 (en) | 1997-09-24 | 2003-04-29 | The Regents Of The University Of California | Lentiviral nucleic acids and uses thereof |
US6790657B1 (en) | 1999-01-07 | 2004-09-14 | The United States Of America As Represented By The Department Of Health And Human Services | Lentivirus vector system |
US7226780B2 (en) | 1999-01-07 | 2007-06-05 | United States Of America As Represented By The Secretary Of The Department Of Health And Human Resources | Lentivirus vector system |
WO2013037841A3 (en) * | 2011-09-12 | 2013-05-23 | Centre National De La Recherche Scientifique (C.N.R.S) | Chimeric non-integrating lentiviral genomes as vaccines against hiv-1 |
US9879230B2 (en) | 2011-09-12 | 2018-01-30 | Centre National De La Recherche Scientifique (C.N.R.S) | Chimeric non-integrating lentiviral genomes as vaccines against HIV-1 |
Also Published As
Publication number | Publication date |
---|---|
EP0752000A1 (en) | 1997-01-08 |
JPH09510361A (en) | 1997-10-21 |
WO1995025806A3 (en) | 1996-01-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Laughrea et al. | Mutations in the kissing-loop hairpin of human immunodeficiency virus type 1 reduce viral infectivity as well as genomic RNA packaging and dimerization | |
Saltarelli et al. | Nucleotide sequence and transcriptional analysis of molecular clones of CAEV which generate infectious virus | |
Coffin | Genetic diversity and evolution of retroviruses | |
US5886166A (en) | Retroviral vector, a replication system for said vector and avian or mammalian cells transfected with said vector | |
US6114141A (en) | Methods to express genes from viral vectors | |
Diener | Origin and evolution of viroids and viroid-like satellite RNAs | |
JP2007014341A (en) | Conditionally replicating viral vectors and their use | |
EP1192265A1 (en) | Anti-viral vectors | |
Russell et al. | Effects of a Single Amino Acid Substitution within thep2 Region of Human Immunodeficiency Virus Type 1 on Packagingof Spliced ViralRNA | |
CN101353671B (en) | Preparation and use of recombinant virus vector gene A3G | |
US10434187B2 (en) | Use of microRNAs to control virus helper nucleic acids | |
EP0752000A1 (en) | Packaging-deficient lentiviruses | |
EP1115879B1 (en) | Retroviral particles protected against complement mediated destruction | |
Robson et al. | Selection of optimal polypurine tract region sequences during Moloney murine leukemia virus replication | |
WO2012118092A1 (en) | Fusion protein | |
Rong et al. | Hydrophobic amino acids in the human immunodeficiency virus type 1 p2 and nucleocapsid proteins can contribute to the rescue of deleted viral RNA packaging signals | |
US20120034693A1 (en) | Recombinant vector and use in gene therapy | |
US5753490A (en) | Recombinant HIV and modified packaging cells and method for treating acquired immune deficiency syndrome | |
AU2004202502B2 (en) | Retroviral particles protected against complement mediated destruction | |
JP2001520017A (en) | Inhibition of human immunodeficiency virus (HIV-1) replication | |
US6063374A (en) | Recombinant HIV and modified packaging cells and method for using | |
WO1999000490A2 (en) | Attenuated human immunodeficiency virus vaccine | |
EP0763123A1 (en) | Safe retroviral vectors, their production and use | |
Ly | Retroviral genomic RNA dimerization: Structural and functional characterizations | |
Carmo | Gammaretroviral and Lentiviral Vectors for Gene Therapy Stability and Inactivation Mechanisms |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A2 Designated state(s): JP US |
|
AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
AK | Designated states |
Kind code of ref document: A3 Designated state(s): JP US |
|
AL | Designated countries for regional patents |
Kind code of ref document: A3 Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1995912349 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 1995912349 Country of ref document: EP |
|
ENP | Entry into the national phase in: |
Ref country code: US Ref document number: 1997 522292 Date of ref document: 19970205 Kind code of ref document: A Format of ref document f/p: F |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 1995912349 Country of ref document: EP |