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

使用带有锌指核酸酶的 ssDNA 寡核苷酸进行高频基因组编辑

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作者:Fuqiang Chen, Shondra M Pruett-Miller, Yuping Huang, Monika Gjoka, Katarzyna Duda, Jack Taunton, Trevor N Collingwood, Morten Frodin, Gregory D Davis

Abstract

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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