WO2004046335A3 - A method for cloning of a rare, specifically mutated cell - Google Patents

A method for cloning of a rare, specifically mutated cell Download PDF

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Publication number
WO2004046335A3
WO2004046335A3 PCT/US2003/036882 US0336882W WO2004046335A3 WO 2004046335 A3 WO2004046335 A3 WO 2004046335A3 US 0336882 W US0336882 W US 0336882W WO 2004046335 A3 WO2004046335 A3 WO 2004046335A3
Authority
WO
WIPO (PCT)
Prior art keywords
rare
alterations
cloning
pcr
alteration
Prior art date
Application number
PCT/US2003/036882
Other languages
French (fr)
Other versions
WO2004046335A2 (en
Inventor
Richard Metz
Mike Dicola
Michael R Blaese
Original Assignee
Richard Metz
Mike Dicola
Michael R Blaese
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 Richard Metz, Mike Dicola, Michael R Blaese filed Critical Richard Metz
Priority to AU2003291073A priority Critical patent/AU2003291073A1/en
Publication of WO2004046335A2 publication Critical patent/WO2004046335A2/en
Publication of WO2004046335A3 publication Critical patent/WO2004046335A3/en

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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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries

Landscapes

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

Abstract

The invention concerns a new method of detecting a rare product of a directed genetic alteration of a cultured cell. The method is applicable to any method of making the alteration. The method comprises sequentially using polymerase chain reaction (PCR) using an allele specific primer to preferentially amplify sequences containing one of the two linked alterations, coupled with a second technique that detects the second change in the PCR product. The second method may comprise restriction digestion, conventional DNA sequencing or pyro-sequencing. Experiments indicate that alterations as rare as one correctly altered copy in 10,000 cells can be detected using the methods provided.
PCT/US2003/036882 2002-11-18 2003-11-18 A method for cloning of a rare, specifically mutated cell WO2004046335A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2003291073A AU2003291073A1 (en) 2002-11-18 2003-11-18 A method for cloning of a rare, specifically mutated cell

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US10/298,850 2002-11-18
US10/298,850 US6929917B2 (en) 2002-11-18 2002-11-18 Method for cloning of a rare, specifically mutated cell

Publications (2)

Publication Number Publication Date
WO2004046335A2 WO2004046335A2 (en) 2004-06-03
WO2004046335A3 true WO2004046335A3 (en) 2009-06-18

Family

ID=32297551

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2003/036882 WO2004046335A2 (en) 2002-11-18 2003-11-18 A method for cloning of a rare, specifically mutated cell

Country Status (3)

Country Link
US (1) US6929917B2 (en)
AU (1) AU2003291073A1 (en)
WO (1) WO2004046335A2 (en)

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6010908A (en) * 1992-08-21 2000-01-04 The Regents Of The University Of California Gene therapy by small fragment homologous replacement
GB9620209D0 (en) * 1996-09-27 1996-11-13 Cemu Bioteknik Ab Method of sequencing DNA
GB9711015D0 (en) 1997-05-28 1997-07-23 Zeneca Ltd Improvements in or relating to organic compounds
CA2298886A1 (en) 1997-08-05 1999-02-18 Kimeragen, Inc. The use of mixed duplex oligonucleotides to effect localized genetic changes in plants
US6271360B1 (en) 1999-08-27 2001-08-07 Valigen (Us), Inc. Single-stranded oligodeoxynucleotide mutational vectors
WO2002026967A2 (en) * 2000-09-25 2002-04-04 Thomas Jefferson University Targeted gene correction by single-stranded oligodeoxynucleotides

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ALEXEEV, ET AL: "LOCALIZED IN VIVO GENOTYPIC PHENOTYPIC CORRECTION OF THE ALBINO MUTATION IN SKIN BY RNA-DNA OLIGONUCLEOTIDE", NATURE BIOTECH, vol. 18, January 2000 (2000-01-01), pages 43 - 47, XP002222806 *
BARTLETT R J, ET AL: "IN VIVO TARGETED REPAIR OF A POINT MUTATION IN THE CANINE DYSTROPHIN GENE BY A CHIMERIC RNA/DNA OLIGONUCLEOTIDE", NATURE BIOTECHNOLOGY, vol. 18, June 2000 (2000-06-01), pages 615 - 622, XP002977240 *
COLE-STRAUSS, ET AL: "TARGETED GENE REPAIR DIRECTED BY THE CHIMERIC RNA/DNA OLIGONUCLEOTIDE IN A MAMMALIAN CELL-FREE EXTRACT", NUCLEIC ACIDS RESEARCH, vol. 27, no. 5, May 1999 (1999-05-01), pages 1323 - 1330, XP002151268 *
YOON, ET AL: "TARGETED GENE CORRECTION OF EPISOMAL DNA IN MAMMALIAN CELLS MEDIATED BY A CHEMERIC RNA/DNA OLIGONUCLEOTIDE", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCE OF USA, vol. 93, March 1996 (1996-03-01), pages 2071 - 2076, XP002156595 *

Also Published As

Publication number Publication date
AU2003291073A8 (en) 2009-07-30
US20040096832A1 (en) 2004-05-20
AU2003291073A1 (en) 2004-06-15
US6929917B2 (en) 2005-08-16
WO2004046335A2 (en) 2004-06-03

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