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WO2010113031A3 - Method of altering nucleic acids - Google Patents

Method of altering nucleic acids Download PDF

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Publication number
WO2010113031A3
WO2010113031A3 PCT/IB2010/000893 IB2010000893W WO2010113031A3 WO 2010113031 A3 WO2010113031 A3 WO 2010113031A3 IB 2010000893 W IB2010000893 W IB 2010000893W WO 2010113031 A3 WO2010113031 A3 WO 2010113031A3
Authority
WO
WIPO (PCT)
Prior art keywords
nucleic acid
sequence
type iis
acid molecule
homology
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/IB2010/000893
Other languages
French (fr)
Other versions
WO2010113031A2 (en
Inventor
Adrian Francis Stewart
Youming Zhang
Marcello Maresca
Harald Kranz
Stephan Noll
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.)
Gene Bridges GmbH
Original Assignee
Gene Bridges GmbH
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 Gene Bridges GmbH filed Critical Gene Bridges GmbH
Publication of WO2010113031A2 publication Critical patent/WO2010113031A2/en
Publication of WO2010113031A3 publication Critical patent/WO2010113031A3/en
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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/102—Mutagenizing nucleic acids
    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1082—Preparation or screening gene libraries by chromosomal integration of polynucleotide sequences, HR-, site-specific-recombination, transposons, 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
    • 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
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
    • C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
    • 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
    • C12N2800/80—Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Virology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

The invention provides a method for altering the sequence of a target nucleic acid molecule, said method comprising the steps of: a) introducing a nucleic acid fragment into the target nucleic acid molecule by homologous recombination, wherein the nucleic acid fragment comprises: i) a first region of homology to the target nucleic acid molecule; ii) a first recognition sequence for a Type IIS nuclease; iii) a selectable marker; iv) a second recognition sequence for a Type IIS nuclease; v) a second region of homology to the target nucleic acid molecule; wherein components i) to v) are ordered from 5' to 3'; and a replacement nucleic acid sequence positioned between the first and second regions of homology but flanking the recognition sequences for the Type IIS nucleases; b) selecting for the incorporation of the replacement nucleic acid using the selectable marker, and c) cleaving the product of step b) with Type IIS nucleases such that the selectable marker is excised, to produce a linear target fragment including the desired replacement sequence.
PCT/IB2010/000893 2009-03-30 2010-03-30 Method of altering nucleic acids Ceased WO2010113031A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0905458A GB0905458D0 (en) 2009-03-30 2009-03-30 Method of altering nucleic acids
GB0905458.6 2009-03-30

Publications (2)

Publication Number Publication Date
WO2010113031A2 WO2010113031A2 (en) 2010-10-07
WO2010113031A3 true WO2010113031A3 (en) 2011-01-27

Family

ID=40671958

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2010/000893 Ceased WO2010113031A2 (en) 2009-03-30 2010-03-30 Method of altering nucleic acids

Country Status (2)

Country Link
GB (1) GB0905458D0 (en)
WO (1) WO2010113031A2 (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2011215557B2 (en) 2010-02-09 2016-03-10 Sangamo Therapeutics, Inc. Targeted genomic modification with partially single-stranded donor molecules
DK2831238T3 (en) 2012-03-27 2018-04-03 Dsm Ip Assets Bv CLONING PROCEDURE
WO2023107713A2 (en) * 2021-12-09 2023-06-15 Bonadea Diagnostics, Llc Sequence conversion reaction

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
CHING YICK PANG ET AL: "Analysis of the specificity of the AMP-activated protein kinase by site-directed mutagenesis of bacterially expressed 3-hydroxy 3-methylglutaryl-CoA reductase, using a single primer variant of the unique-site-elimination method", EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 237, no. 3, 1996, pages 800 - 808, XP002604746, ISSN: 0014-2956 *
DENG W P ET AL: "Site-directed mutagenesis of virtually any plasmid by eliminating a unique site", ANALYTICAL BIOCHEMISTRY, vol. 200, no. 1, 1 January 1992 (1992-01-01), ACADEMIC PRESS INC, NEW YORK, pages 81 - 88, XP024819821, ISSN: 0003-2697, [retrieved on 19920101], DOI: 10.1016/0003-2697(92)90280-K *
LI ET AL: "Site-directed mutagenesis by combination of homologous recombination and DpnI digestion of the plasmid template in Escherichia coli", ANALYTICAL BIOCHEMISTRY, vol. 373, no. 2, 30 October 2007 (2007-10-30), ACADEMIC PRESS INC, NEW YORK, pages 389 - 391, XP022411123, ISSN: 0003-2697, DOI: 10.1016/J.AB.2007.10.034 *
MUYRERS J P P ET AL: "Techniques: Recombinogenic engineering: New options for cloning and manipulating DNA", TRENDS IN BIOCHEMICAL SCIENCES, vol. 26, no. 5, 1 May 2001 (2001-05-01), ELSEVIER, HAYWARDS, GB, pages 325 - 331, XP002227320, ISSN: 0968-0004, DOI: 10.1016/S0968-0004(00)01757-6 *
NOLL STEPHAN ET AL: "Site-directed mutagenesis of multi-copy-number plasmids: Red/ET recombination and unique restriction site elimination", BIOTECHNIQUES, vol. 46, no. 7, June 2009 (2009-06-01), XP002604747, ISSN: 0736-6205 *
POSFAI G ET AL: "Markerless gene replacement in escherichia coli stimulated by a double-strand break in the chromosome", NUCLEIC ACIDS RESEARCH, vol. 27, no. 22, 15 November 1999 (1999-11-15), OXFORD UNIVERSITY PRESS, SURREY, GB, pages 4409 - 4415, XP002963851, ISSN: 0305-1048, DOI: 10.1093/NAR/27.22.4409 *
WALKER M ET AL: "PCR-based gene disruption and recombinatory marker excision to produce modified industrial Saccharomyces cerevisiae without added sequences", JOURNAL OF MICROBIOLOGICAL METHODS, vol. 63, no. 2, 1 November 2005 (2005-11-01), ELSEVIER, AMSTERDAM, NL, pages 193 - 204, XP025258997, ISSN: 0167-7012, [retrieved on 20051101], DOI: 10.1016/J.MIMET.2005.03.015 *
ZHANG Y ET AL: "A new logic for DNA engineering using recombination in Escherichia coli", NATURE GENETICS, vol. 20, no. 2, 1 October 1998 (1998-10-01), NATURE PUBLISHING GROUP, NEW YORK, US, pages 123 - 128, XP002225129, ISSN: 1061-4036, DOI: 10.1038/2417 *
ZHANG YOUMING ET AL: "Phage annealing proteins promote oligonucleotide-directed mutagenesis in Escherichia coli and mouse ES cells", BMC MOLECULAR BIOLOGY, vol. 4, no. 1, 16 January 2003 (2003-01-16), BIOMED CENTRAL LTD, GB, pages 1, XP021014950, ISSN: 1471-2199, DOI: 10.1186/1471-2199-4-1 *

Also Published As

Publication number Publication date
GB0905458D0 (en) 2009-05-13
WO2010113031A2 (en) 2010-10-07

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