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

High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases.
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DOI:
10.1038/nmeth.1653
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发表时间:
2011-07-17
期刊:
影响因子:
48
通讯作者:
Davis, Gregory D.
Davis, Gregory D.
中科院分区:
生物学1区
文献类型:
--
作者:
Chen, Fuqiang;Pruett-Miller, Shondra M.;Huang, Yuping;Gjoka, Monika;Duda, Katarzyna;Taunton, Jack;Collingwood, Trevor N.;Frodin, Morten;Davis, Gregory D.

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锌指核酸酶 (ZFN) 能够在多种细胞类型和生物体中实现高效的基因靶向。在这里,我们描述了使用简单的 ssDNA 寡核苷酸与 ZFN 串联的方法,以有效地产生具有三种不同遗传结果的人类细胞系:(i)靶向点突变,(ii)高达 100 kb 的靶向基因组删除和(iii)伴随大基因组删除的小遗传元件的靶向插入。
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.
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