Easy quantification of template-directed CRISPR/Cas9 editing

Easy quantification of template-directed CRISPR/Cas9 editing
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DOI:
10.1093/nar/gky164
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发表时间:
2018-06-01
影响因子:
14.9
通讯作者:
van Steensel, Bas
van Steensel, Bas
中科院分区:
生物学2区
文献类型:
--
作者:
Brinkman, Eva K.;Kousholt, Arne N.;van Steensel, Bas

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模板指导的CRISPR/Cas9编辑是在基因组中引入细微突变的强大工具。然而,在靶位点掺入所需突变的成功率难以预测,因此必须凭经验确定。在这里,我们采用了广泛使用的TIDE方法来定量模板化编辑事件,包括点突变。由此产生的TIDER方法是用于测试和优化模板指导的基因组编辑策略的快速,廉价和可访问的工具。
Template-directed CRISPR/Cas9 editing is a powerful tool for introducing subtle mutations in genomes. However, the success rate of incorporation of the desired mutations at the target site is difficult to predict and therefore must be empirically determined. Here, we adapted the widely used TIDE method for quantification of templated editing events, including point mutations. The resulting TIDER method is a rapid, cheap and accessible tool for testing and optimization of template-directed genome editing strategies.