WO2009146668A3 - Regulatory gene element comprising promotor/enhancer protected from dna methylation and gene expression silencing - Google Patents

Regulatory gene element comprising promotor/enhancer protected from dna methylation and gene expression silencing Download PDF

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Publication number
WO2009146668A3
WO2009146668A3 PCT/CZ2009/000079 CZ2009000079W WO2009146668A3 WO 2009146668 A3 WO2009146668 A3 WO 2009146668A3 CZ 2009000079 W CZ2009000079 W CZ 2009000079W WO 2009146668 A3 WO2009146668 A3 WO 2009146668A3
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WO
WIPO (PCT)
Prior art keywords
gene
sequence
enhancer
ltr
methylation
Prior art date
Application number
PCT/CZ2009/000079
Other languages
French (fr)
Other versions
WO2009146668A2 (en
Inventor
Jiri Hejnar
Filip Senigl
Original Assignee
Ustav Molekulami Genetiky Av Cr, V.V.I.
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 Ustav Molekulami Genetiky Av Cr, V.V.I. filed Critical Ustav Molekulami Genetiky Av Cr, V.V.I.
Publication of WO2009146668A2 publication Critical patent/WO2009146668A2/en
Publication of WO2009146668A3 publication Critical patent/WO2009146668A3/en

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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
    • 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
    • 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
    • C12N2740/11011—Alpharetrovirus, e.g. avian leucosis virus
    • C12N2740/11041—Use of virus, viral particle or viral elements as a vector
    • C12N2740/11043—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
    • C12N2740/13011—Gammaretrovirus, e.g. murine leukeamia virus
    • C12N2740/13041—Use of virus, viral particle or viral elements as a vector
    • C12N2740/13043—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
    • C12N2830/00—Vector systems having a special element relevant for transcription
    • C12N2830/46—Vector systems having a special element relevant for transcription elements influencing chromatin structure, e.g. scaffold/matrix attachment region, methylation free island

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Virology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The invention relates to the field of molecular biology, virology, and gene therapy where various vectors, particularly viral and retroviral vectors, might be used for gene transfer either into the cultivated cells in vitro or into living organisms. Specifically, the invention provides solution to the problem of silenced gene expression and methylation of the vector DNA in eukaryotic cells, particularly in mammalian cells. This solution consists in the use of a specific regulatory gene element containing a promoter/enhancer based on the retroviral long terminal repeat (LTR) modified by insertion of at least one internal element (IE) from the CpG island. The LTR modified in this way ensures strong and long-term expression of the integrated vector together with the transduced gene of interest (transgene) and, at he same time, its efficient protection from transcriptional silencing and gradual methylation, even in human cells. Particularly, the invention discloses a regulatory gene element comprising the enhancer/promoter protected from DNA methylation and from silencing of gene expression, wherein said element comprises a modified retroviral sequence LTR with at least one IE sequence inserted in any orientation into any position, wherein the IE sequence comprises SEQUENCE ID No. 2. or any equivalent sequence that fulfils the same function.
PCT/CZ2009/000079 2008-06-03 2009-06-03 Regulatory gene element comprising promotor/enhancer protected from dna methylation and gene expression silencing WO2009146668A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CZ20080343A CZ2008343A3 (en) 2008-06-03 2008-06-03 Regulating gene element comprising promoter/amplifier protected from DNA methylation and gene expression suppression
CZPV2008-343 2008-06-03

Publications (2)

Publication Number Publication Date
WO2009146668A2 WO2009146668A2 (en) 2009-12-10
WO2009146668A3 true WO2009146668A3 (en) 2010-01-28

Family

ID=40590041

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CZ2009/000079 WO2009146668A2 (en) 2008-06-03 2009-06-03 Regulatory gene element comprising promotor/enhancer protected from dna methylation and gene expression silencing

Country Status (2)

Country Link
CZ (1) CZ2008343A3 (en)
WO (1) WO2009146668A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SG191246A1 (en) * 2010-12-24 2013-07-31 Agency Science Tech & Res Modified human cmv promoters that are resistant to gene silencing

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002094989A2 (en) * 2001-05-18 2002-11-28 The Scripps Research Institute Retroviral vectors and methods of using same

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002094989A2 (en) * 2001-05-18 2002-11-28 The Scripps Research Institute Retroviral vectors and methods of using same

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DATABASE GENESEQ 28 May 2003 (2003-05-28), XP002556313, retrieved from EBI Database accession no. AAD53297 *
HEJNAR J ET AL: "CpG island protects Rous sarcoma virus-derived vectors integrated into nonpermissive cells from DNA methylation and transcriptional suppression.", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 16 JAN 2001, vol. 98, no. 2, 16 January 2001 (2001-01-16), pages 565 - 569, XP002556300, ISSN: 0027-8424 *
SENIGL FILIP ET AL: "The core element of a CpG island protects avian sarcoma and leukosis virus-derived vectors from transcriptional silencing.", JOURNAL OF VIROLOGY AUG 2008, vol. 82, no. 16, August 2008 (2008-08-01), pages 7818 - 7827, XP002556302, ISSN: 1098-5514 *
SIEGFRIED Z ET AL: "DNA methylation represses transcription in vivo.", NATURE GENETICS JUN 1999, vol. 22, no. 2, June 1999 (1999-06-01), pages 203 - 206, XP002556301, ISSN: 1061-4036 *

Also Published As

Publication number Publication date
CZ300377B6 (en) 2009-05-06
CZ2008343A3 (en) 2009-05-06
WO2009146668A2 (en) 2009-12-10

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