High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases

被引：340

作者：

Chen, Fuqiang ^{[1
]}

Pruett-Miller, Shondra M. ^{[1
]}

Huang, Yuping ^{[1
]}

Gjoka, Monika ^{[1
]}

Duda, Katarzyna ^{[2
,3
]}

Taunton, Jack ^{[4
]}

Collingwood, Trevor N. ^{[1
]}

Frodin, Morten ^{[2
,3
]}

Davis, Gregory D. ^{[1
]}

机构：

[1] Sigma Aldrich Biotechnol, St Louis, MO USA

[2] Univ Copenhagen, Biotech Res & Innovat Ctr, Copenhagen, Denmark

[3] Univ Copenhagen, Ctr Epigenet, Copenhagen, Denmark

[4] Univ Calif San Francisco, Howard Hughes Med Inst, Dept Cellular & Mol Pharmacol, San Francisco, CA USA

来源：

NATURE METHODS | 2011年 / 8卷 / 09期

基金：

美国国家卫生研究院; 新加坡国家研究基金会;

关键词：

DOUBLE-STRAND BREAKS; MAMMALIAN-CELLS; GENE KNOCKOUT; REPAIR; PATHWAY; LOCUS;

D O I：

10.1038/nmeth.1653

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point mutation, (ii) targeted genomic deletion of up to 100 kb and (iii) targeted insertion of small genetic elements concomitant with large genomic deletions.

引用

收藏

页码：753 / U96

页数：5

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