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WO2000065076A3 - Synthetic gene for expressing active retroviral protein in eukaryotes - Google Patents

Synthetic gene for expressing active retroviral protein in eukaryotes Download PDF

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Publication number
WO2000065076A3
WO2000065076A3 PCT/EP2000/003765 EP0003765W WO0065076A3 WO 2000065076 A3 WO2000065076 A3 WO 2000065076A3 EP 0003765 W EP0003765 W EP 0003765W WO 0065076 A3 WO0065076 A3 WO 0065076A3
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WO
WIPO (PCT)
Prior art keywords
protein
enzyme
retroviral
gene
integrase
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/EP2000/003765
Other languages
French (fr)
Other versions
WO2000065076A2 (en
Inventor
Zeger Debyser
Clercq Erik De
Peter Cherepanov
Wim Pluymers
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.)
Katholieke Universiteit Leuven
Original Assignee
Katholieke Universiteit Leuven
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 Katholieke Universiteit Leuven filed Critical Katholieke Universiteit Leuven
Priority to JP2000614410A priority Critical patent/JP2002542791A/en
Priority to CA002369058A priority patent/CA2369058A1/en
Priority to IL14609000A priority patent/IL146090A0/en
Priority to AU45579/00A priority patent/AU778106B2/en
Priority to BR0010077-3A priority patent/BR0010077A/en
Priority to EP00927069A priority patent/EP1183381A2/en
Publication of WO2000065076A2 publication Critical patent/WO2000065076A2/en
Priority to NO20015168A priority patent/NO20015168L/en
Anticipated expiration legal-status Critical
Publication of WO2000065076A3 publication Critical patent/WO2000065076A3/en
Ceased legal-status Critical Current

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    • 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
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • 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
    • 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
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16041—Use of virus, viral particle or viral elements as a vector
    • C12N2740/16043—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
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16051—Methods of production or purification of viral material
    • C12N2740/16052—Methods of production or purification of viral material relating to complementing cells and packaging systems for producing virus or viral particles
    • 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
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/10—Plasmid DNA
    • C12N2800/108—Plasmid DNA episomal vectors
    • 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
    • C12N2840/00—Vectors comprising a special translation-regulating system
    • C12N2840/20—Vectors comprising a special translation-regulating system translation of more than one cistron
    • C12N2840/203—Vectors comprising a special translation-regulating system translation of more than one cistron having an IRES

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Virology (AREA)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The present invention features a synthetic gene or region of a gene which has an amended codon usage compared with the wild-type gene and which is for the high level expression of a retroviral protein in eukaryotic cells, the expressed retroviral protein having enzymatic activity in the eukaryotic cell. In addition, the invention features a synthetic gene or region of a gene encoding a retroviral enzyme or part of a retroviral enzyme normally expressed in a mammalian or other eukaryotic cell wherein at least one non-preferred codon in the wild-type gene encoding the enzyme has been replaced by a preferred codon encoding the same amino acid. The retroviral protein may be a protease, reverse transcriptase, integrase protein or a polyprotein gag-pol precursor thereof. In one embodiment the retroviral protein with enzymatic activity is a lentiviral protein. In other embodiments the enzymatically active protein is a pol enzyme. In more preferred embodiments, the enzymatically active protein is a lentiviral integrase. In an even more preferred embodiment the enzyme is an HIV enzyme. In more preferred embodiments the enzymatically active protein is HIV integrase. The present invention also includes a detection method for intracellular integrase using a promoterless reporter gene.
PCT/EP2000/003765 1999-04-26 2000-04-26 Synthetic gene for expressing active retroviral protein in eukaryotes Ceased WO2000065076A2 (en)

Priority Applications (7)

Application Number Priority Date Filing Date Title
JP2000614410A JP2002542791A (en) 1999-04-26 2000-04-26 Synthetic genes for expressing active retroviral proteins in eukaryotes
CA002369058A CA2369058A1 (en) 1999-04-26 2000-04-26 Synthetic gene for expressing active retroviral protein in eukaryotes
IL14609000A IL146090A0 (en) 1999-04-26 2000-04-26 A synthetic gene for expression of a retroviral protein with wild type activity in eukaryotic cells
AU45579/00A AU778106B2 (en) 1999-04-26 2000-04-26 Synthetic gene for expressing active retroviral protein in eukaryotes
BR0010077-3A BR0010077A (en) 1999-04-26 2000-04-26 Synthetic gene for expression of an active retroviral protein in eukaryotes
EP00927069A EP1183381A2 (en) 1999-04-26 2000-04-26 Synthetic gene for expressing active retroviral protein in eukaryotes
NO20015168A NO20015168L (en) 1999-04-26 2001-10-23 Synthetic gene for expression of active retroviral protein in eukaryotes

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP99201306 1999-04-26
EP99201306.0 1999-04-26
EP00200171 2000-01-18
EP00200171.7 2000-01-18

Publications (2)

Publication Number Publication Date
WO2000065076A2 WO2000065076A2 (en) 2000-11-02
WO2000065076A3 true WO2000065076A3 (en) 2001-12-13

Family

ID=26071747

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2000/003765 Ceased WO2000065076A2 (en) 1999-04-26 2000-04-26 Synthetic gene for expressing active retroviral protein in eukaryotes

Country Status (11)

Country Link
EP (1) EP1183381A2 (en)
JP (1) JP2002542791A (en)
AU (1) AU778106B2 (en)
BR (1) BR0010077A (en)
CA (1) CA2369058A1 (en)
HU (1) HUP0201019A2 (en)
IL (1) IL146090A0 (en)
NO (1) NO20015168L (en)
PL (1) PL351988A1 (en)
RU (1) RU2001131728A (en)
WO (1) WO2000065076A2 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7211659B2 (en) 2001-07-05 2007-05-01 Chiron Corporation Polynucleotides encoding antigenic HIV type C polypeptides, polypeptides and uses thereof
US7282364B2 (en) 2001-08-31 2007-10-16 Novartis Vaccines And Diagnostics, Inc. Polynucleotides encoding antigenic HIV type B polypeptides, polypeptides and uses thereof
US7348177B2 (en) 1998-12-31 2008-03-25 Novartis Vaccines And Diagnostics, Inc. Expression of HIV polypeptides and production of virus-like particles

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9803351D0 (en) 1998-02-17 1998-04-15 Oxford Biomedica Ltd Anti-viral vectors
CA2358385C (en) 1998-12-31 2013-08-06 Chiron Corporation Polynucleotides encoding antigenic hiv type c polypeptides, polypeptides and uses thereof
JP4627950B2 (en) * 1999-12-23 2011-02-09 アメリカ合衆国 Molecular clone with mutant HIV gag / pol, SIV gag and SIV env genes
US6656706B2 (en) * 1999-12-23 2003-12-02 The United States Of America As Represented By The Department Of Health And Human Services Molecular clones with mutated HIV gag/pol, SIV gag and SIV env genes
AUPQ580300A0 (en) * 2000-02-23 2000-03-16 National Cancer Centre Of Singapore Pte Ltd Genetic analysis
GB0009760D0 (en) * 2000-04-19 2000-06-07 Oxford Biomedica Ltd Method
US6835568B2 (en) 2001-10-30 2004-12-28 Virxsys Corporation Regulated nucleic acid expression system
JP4385135B2 (en) * 2003-05-06 2009-12-16 独立行政法人産業技術総合研究所 Multi-gene transcriptional activity measurement system
US8106176B2 (en) 2005-10-07 2012-01-31 Instituto di Richerche di Biologia Molecolare P. Angeletti SpA Matrix metalloproteinase 11 vaccine
JP6824594B2 (en) 2014-09-11 2021-02-03 Jnc株式会社 How to design synthetic genes

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US5434065A (en) * 1993-05-06 1995-07-18 President And Fellows Of Harvard College In vivo selection of microbial virulence genes
US5468629A (en) * 1993-04-13 1995-11-21 Calhoun; Cornelia Method of promoting in vitro homologous recombination transfection in mammalian cells using the RecA protein
WO1997036481A1 (en) * 1996-04-02 1997-10-09 The Regents Of The University Of California Design of novel highly efficient hiv based packaging systems for gene therapy
WO1998012207A1 (en) * 1996-09-20 1998-03-26 The General Hospital Corporation High level expression of proteins
WO1998034640A2 (en) * 1997-02-07 1998-08-13 Merck & Co., Inc. Synthetic hiv gag genes
US5811270A (en) * 1994-05-20 1998-09-22 Grandgenett; Duane P. In vitro method for concerted integration of donor DNA molecules using retroviral integrase proteins
WO2000039302A2 (en) * 1998-12-31 2000-07-06 Chiron Corporation Improved expression of hiv polypeptides and production of virus-like particles

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2358385C (en) * 1998-12-31 2013-08-06 Chiron Corporation Polynucleotides encoding antigenic hiv type c polypeptides, polypeptides and uses thereof

Patent Citations (7)

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US5468629A (en) * 1993-04-13 1995-11-21 Calhoun; Cornelia Method of promoting in vitro homologous recombination transfection in mammalian cells using the RecA protein
US5434065A (en) * 1993-05-06 1995-07-18 President And Fellows Of Harvard College In vivo selection of microbial virulence genes
US5811270A (en) * 1994-05-20 1998-09-22 Grandgenett; Duane P. In vitro method for concerted integration of donor DNA molecules using retroviral integrase proteins
WO1997036481A1 (en) * 1996-04-02 1997-10-09 The Regents Of The University Of California Design of novel highly efficient hiv based packaging systems for gene therapy
WO1998012207A1 (en) * 1996-09-20 1998-03-26 The General Hospital Corporation High level expression of proteins
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WO2000039302A2 (en) * 1998-12-31 2000-07-06 Chiron Corporation Improved expression of hiv polypeptides and production of virus-like particles

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7348177B2 (en) 1998-12-31 2008-03-25 Novartis Vaccines And Diagnostics, Inc. Expression of HIV polypeptides and production of virus-like particles
US7211659B2 (en) 2001-07-05 2007-05-01 Chiron Corporation Polynucleotides encoding antigenic HIV type C polypeptides, polypeptides and uses thereof
US9598469B2 (en) 2001-07-05 2017-03-21 Novartis Vaccines And Diagnostics, Inc. HIV-1 south african subtype C env proteins
US7282364B2 (en) 2001-08-31 2007-10-16 Novartis Vaccines And Diagnostics, Inc. Polynucleotides encoding antigenic HIV type B polypeptides, polypeptides and uses thereof

Also Published As

Publication number Publication date
IL146090A0 (en) 2002-07-25
WO2000065076A2 (en) 2000-11-02
CA2369058A1 (en) 2000-11-02
AU4557900A (en) 2000-11-10
AU778106B2 (en) 2004-11-18
NO20015168L (en) 2001-12-14
PL351988A1 (en) 2003-07-14
NO20015168D0 (en) 2001-10-23
EP1183381A2 (en) 2002-03-06
JP2002542791A (en) 2002-12-17
RU2001131728A (en) 2004-02-20
BR0010077A (en) 2002-01-15
HUP0201019A2 (en) 2002-08-28

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