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

被引:125
作者
Kohli, M
Rago, C
Lengauer, C
Kinzler, KW
Vogelstein, B [1 ]
机构
[1] Johns Hopkins Univ, Inst Med, Howard Hughes Med Inst, Baltimore, MD 21231 USA
[2] Johns Hopkins Univ, Inst Med, Sidney Kimmel Comprehens Canc Ctr, Baltimore, MD 21231 USA
[3] Johns Hopkins Univ, Inst Med, Cellular & Mol Med Program, Baltimore, MD 21231 USA
关键词
D O I
10.1093/nar/gnh009
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
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页数:8
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