Rapid Screening of CRISPR/Cas9-Induced Mutants Using the ACT-PCR Method

文献类型： 外文期刊

第一作者： Wang, Chun

作者： Wang, Chun;Wang, Kejian

作者机构： Chinese Acad Agr Sci, China Natl Rice Res Inst, State Key Lab Rice Biol, Hangzhou, Peoples R China

关键词： ACT-PCR; CRISPR/Cas9; Genome editing; Mutant screening; Rapid; Large-scale; Cost-effective

期刊名称：PLANT GENOME EDITING WITH CRISPR SYSTEMS: METHODS AND PROTOCOLS

ISSN： 1064-3745

年卷期： 2019 年 1917 卷

页码：

收录情况： SCI

摘要： The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system provides a technological breakthrough in targeted mutagenesis. However, a significant amount of time and cost is required to screen for the CRISPR/Cas9-induced mutants from a typically large number of initial samples. Here, we describe a cost-effective and sensitive screening technique based on conventional polymerase chain reaction (PCR), termed "annealing at critical temperature PCR" (ACT-PCR), for identifying mutants. ACT-PCR requires only a single PCR step followed by agarose gel electrophoresis. The simplicity of ACT-PCR makes it particularly suitable for rapid, large-scale screening of CRISPR/Cas9-induced mutants.

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