Facile methods for generating human somatic cell gene knockouts using recombinant adeno-associated viruses.

Manu Kohli, Carlo Rago, Christoph Lengauer, Kenneth W Kinzler, Bert Vogelstein

Research output: Contribution to journalArticle

Abstract

Emerging evidence suggests that recombinant adeno-associated viral (rAAV) vectors can be used for specific gene targeting in human somatic cells. We have developed an rAAV vector construction procedure employing fusion PCR and a single cloning step that considerably simplifies the knockout process. We demonstrate its utility by disrupting genes at specific positions within human colon cancer cells as well as within immortalized normal epithelial cells. This technology should be broadly applicable to in vitro studies that require the manipulation of the human genome.

Original languageEnglish (US)
JournalNucleic Acids Research
Volume32
Issue number1
StatePublished - 2004

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Dependovirus
Gene Knockout Techniques
Gene Targeting
Human Genome
Colonic Neoplasms
Organism Cloning
Epithelial Cells
Technology
Polymerase Chain Reaction
Genes
In Vitro Techniques

ASJC Scopus subject areas

  • Genetics

Cite this

Facile methods for generating human somatic cell gene knockouts using recombinant adeno-associated viruses. / Kohli, Manu; Rago, Carlo; Lengauer, Christoph; Kinzler, Kenneth W; Vogelstein, Bert.

In: Nucleic Acids Research, Vol. 32, No. 1, 2004.

Research output: Contribution to journalArticle

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