WO2009048024A1 - 目的遺伝子を染色体外で高度に増幅させるためのベクターおよびその利用 - Google Patents

目的遺伝子を染色体外で高度に増幅させるためのベクターおよびその利用 Download PDF

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Publication number
WO2009048024A1
WO2009048024A1 PCT/JP2008/068041 JP2008068041W WO2009048024A1 WO 2009048024 A1 WO2009048024 A1 WO 2009048024A1 JP 2008068041 W JP2008068041 W JP 2008068041W WO 2009048024 A1 WO2009048024 A1 WO 2009048024A1
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WO
WIPO (PCT)
Prior art keywords
target gene
vector
extrachromosomally
amplifying target
highly
Prior art date
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Ceased
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PCT/JP2008/068041
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English (en)
French (fr)
Inventor
Noriaki Shimizu
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Hiroshima University NUC
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Hiroshima University NUC
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Publication date
Application filed by Hiroshima University NUC filed Critical Hiroshima University NUC
Priority to JP2009536987A priority Critical patent/JP5283044B2/ja
Publication of WO2009048024A1 publication Critical patent/WO2009048024A1/ja
Anticipated expiration legal-status Critical
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
    • 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
    • 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
    • C12N2800/00—Nucleic acids 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
    • C12N2820/00—Vectors comprising a special origin of replication system
    • C12N2820/80—Vectors comprising a special origin of replication system from vertebrates
    • C12N2820/85—Vectors comprising a special origin of replication system from vertebrates mammalian

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

Abstract

　本発明は宿主細胞の染色体外で増幅するベクターＤＮＡ、および当該ベクターを用いた目的遺伝子の増幅方法を提供する。本発明にかかるベクターは、真核生物細胞内で機能する哺乳動物複製開始領域、および核マトリックス結合領域を具備するベクターであって、直鎖状であり、かつ少なくとも片側の末端の形状がヘアピン構造であることを特徴としている。本発明のベクターを用いることで、目的遺伝子をＤＭの形態、すなわち染色体外で高度に増幅させることが可能となる。したがって、反復配列依存的な転写抑制を受けることなく、目的遺伝子の増幅コピー数の増加に比例して、目的遺伝子から発現するタンパク質量も増加すると考えられる。
PCT/JP2008/068041 2007-10-09 2008-10-03 目的遺伝子を染色体外で高度に増幅させるためのベクターおよびその利用 Ceased WO2009048024A1 (ja)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2009536987A JP5283044B2 (ja) 2007-10-09 2008-10-03 目的遺伝子を染色体外で高度に増幅させるためのベクターおよびその利用

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2007-263258 2007-10-09
JP2007263258 2007-10-09

Publications (1)

Publication Number Publication Date
WO2009048024A1 true WO2009048024A1 (ja) 2009-04-16

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ID=40549169

Family Applications (1)

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PCT/JP2008/068041 Ceased WO2009048024A1 (ja) 2007-10-09 2008-10-03 目的遺伝子を染色体外で高度に増幅させるためのベクターおよびその利用

Country Status (2)

Country Link
JP (1) JP5283044B2 (ja)
WO (1) WO2009048024A1 (ja)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010110340A1 (ja) 2009-03-27 2010-09-30 国立大学法人広島大学 哺乳動物細胞内で目的遺伝子を増幅し高発現させる方法、および当該方法を実施するために用いられるキット
WO2013146480A1 (ja) * 2012-03-29 2013-10-03 タカラバイオ株式会社 遺伝子導入方法
US9102943B2 (en) 2010-10-05 2015-08-11 Takara Bio Inc. Method for producing virus vector
US9399780B2 (en) 2012-03-22 2016-07-26 Takara Bio Inc. Method for producing viral vector

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP7202573B2 (ja) 2017-11-02 2023-01-12 国立大学法人鳥取大学 哺乳類人工染色体ベクターを利用するタンパク質の高生産方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6451563B1 (en) * 1998-06-15 2002-09-17 Mologen Forschungs-, Entwicklungs- Und Vertriebs Gmbh Method for making linear, covalently closed DNA constructs
JP2003510087A (ja) * 1999-09-27 2003-03-18 ジェネンテック・インコーポレーテッド アポトーシス阻害剤を用いた組換えタンパク質の作成方法
JP2003245083A (ja) * 2002-02-25 2003-09-02 Univ Hiroshima 哺乳動物細胞内で目的遺伝子を高度に増幅させるためのベクターおよび方法
JP2004337066A (ja) * 2003-05-15 2004-12-02 Univ Hiroshima 哺乳動物細胞内にトランスフェクションした遺伝子の安定性を向上させるためのベクターおよび方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6451563B1 (en) * 1998-06-15 2002-09-17 Mologen Forschungs-, Entwicklungs- Und Vertriebs Gmbh Method for making linear, covalently closed DNA constructs
JP2003510087A (ja) * 1999-09-27 2003-03-18 ジェネンテック・インコーポレーテッド アポトーシス阻害剤を用いた組換えタンパク質の作成方法
JP2003245083A (ja) * 2002-02-25 2003-09-02 Univ Hiroshima 哺乳動物細胞内で目的遺伝子を高度に増幅させるためのベクターおよび方法
JP2004337066A (ja) * 2003-05-15 2004-12-02 Univ Hiroshima 哺乳動物細胞内にトランスフェクションした遺伝子の安定性を向上させるためのベクターおよび方法

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
ESCHER G. ET AL.: "Demethylation using the epigenetic modifier, 5-azacytidine, increases the efficiency of transient transfection of macrophages.", JOURNAL OF LIPID RESEARCH, vol. 46, 2005, pages 356 - 365 *
HORIE K. ET AL.: "Gene Targeting by a vector with hairpin-shaped oligonucleotide caps.", BIOCHEMISTRY AND MOLECULAR BIOLOGY INTERNATIONAL, vol. 32, 1994, pages 1041 - 1048 *
SCHAKOWSKI F. ET AL.: "A Novel Minimal-Size Vector (MIDGE) Inproves Transgene Expression in Colon Carcinoma Cells and Avoids Transfection of Undesired DNA.", MOLECULAR THERAPY, vol. 3, 2001, pages 793 - 800 *
SCHAKOWSKI F. ET AL.: "Minimal Size MIDGE Vectors Improve Transgene Expression In Vivo.", IN VIVO, vol. 21, 2 January 2007 (2007-01-02), pages 17 - 23 *
SHIMIZU N. ET AL.: "Amplification of Plasmids Containing a Mammalian Replication Initiation Region Is Mediated by Controllable Conflict between Replication and Transcription.", CANCER RESEARCH, vol. 63, 2003, pages 5281 - 5290 *
SHIMIZU N. ET AL.: "Plasmids with a Mammalian Replication Origin and a Matrix Attachment Region Initiate the Event Similar to Gene Amplification.", CANCER RESEARCH, vol. 61, 2001, pages 6987 - 6990 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010110340A1 (ja) 2009-03-27 2010-09-30 国立大学法人広島大学 哺乳動物細胞内で目的遺伝子を増幅し高発現させる方法、および当該方法を実施するために用いられるキット
US9102943B2 (en) 2010-10-05 2015-08-11 Takara Bio Inc. Method for producing virus vector
US9399780B2 (en) 2012-03-22 2016-07-26 Takara Bio Inc. Method for producing viral vector
WO2013146480A1 (ja) * 2012-03-29 2013-10-03 タカラバイオ株式会社 遺伝子導入方法

Also Published As

Publication number Publication date
JPWO2009048024A1 (ja) 2011-02-17
JP5283044B2 (ja) 2013-09-04

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