Low incidence of off-target mutations in individual CRISPR-Cas9 and TALEN targeted human stem cell clones detected by whole-genome sequencing.
Low incidence of off-target mutations in individual CRISPR-Cas9 and TALEN targeted human stem cell clones detected by whole-genome sequencing.
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DOI：
10.1016/j.stem.2014.04.020
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发表时间：
2014-07-03
期刊：
影响因子：
23.9
通讯作者：
Musunuru, Kiran
中科院分区：
文献类型：
--
作者：
Veres, Adrian;Gosis, Bridget S.;Ding, Qiurong;Collins, Ryan;Ragavendran, Ashok;Brand, Harrison;Erdin, Serkan;Cowan, Chad A.;Talkowski, Michael E.;Musunuru, Kiran
基因组编辑已经吸引了广泛的兴趣，用于使用人类多能干细胞和其他细胞类型生成疾病的细胞模型。CRISPR-Cas系统和TALEN可以在人类细胞中高效靶向所需的基因组位点，但最近的出版物引起了人们对这些工具可能导致脱靶诱变效应的程度的担忧，这些效应可能会混淆疾病建模研究。使用CRISPR-Cas9和TALEN靶向的人多能干细胞克隆，我们以高覆盖率进行了全基因组测序，以评估整个基因组的诱变程度。在这两种类型的克隆中，我们发现归因于核酸酶的脱靶突变非常罕见。从这项分析中，我们认为，虽然某些细胞类型可能存在脱靶突变的风险，但人类多能干细胞中这种效应的发生率可能足够低，不会成为疾病建模和其他应用的重要问题。
Genome editing has attracted wide interest for the generation of cellular models of disease using human pluripotent stem cells and other cell types. CRISPR-Cas systems and TALENs can target desired genomic sites with high efficiency in human cells, but recent publications have led to concern about the extent to which these tools may cause off-target mutagenic effects that could potentially confound disease-modeling studies. Using CRISPR-Cas9 and TALEN targeted human pluripotent stem cell clones, we performed whole-genome sequencing at high coverage to assess the degree of mutagenesis across the entire genome. In both types of clones, we found that off-target mutations attributable to the nucleases were very rare. From this analysis, we suggest that while some cell types may be at risk for off-target mutations, the incidence of such effects in human pluripotent stem cells may be sufficiently low to not be a significant concern for disease modeling and other applications.
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影响因子：
7
作者：
Cho SW;Kim S;Kim Y;Kweon J;Kim HS;Bae S;Kim JS
通讯作者：
Kim JS
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
46.9
作者：
Cho, Seung Woo;Kim, Sojung;Kim, Jin-Soo
通讯作者：
Kim, Jin-Soo
DOI：
10.1126/science.1232033
发表时间：
2013-02-15
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Mali P;Yang L;Esvelt KM;Aach J;Guell M;DiCarlo JE;Norville JE;Church GM
通讯作者：
Church GM
影响因子：
4.3
作者：
Musunuru K
通讯作者：
Musunuru K