Mou Haiwei, Smith Jordan L, Peng Lingtao, Yin Hao, Moore Jill, Zhang Xiao-Ou, Song Chun-Qing, Sheel Ankur, Wu Qiongqiong, Ozata Deniz M, Li Yingxiang, Anderson Daniel G, Emerson Charles P, Sontheimer Erik J, Moore Melissa J, Weng Zhiping, Xue Wen
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02142, USA.
Genome Biol. 2017 Jun 14;18(1):108. doi: 10.1186/s13059-017-1237-8.
CRISPR is widely used to disrupt gene function by inducing small insertions and deletions. Here, we show that some single-guide RNAs (sgRNAs) can induce exon skipping or large genomic deletions that delete exons. For example, CRISPR-mediated editing of β-catenin exon 3, which encodes an autoinhibitory domain, induces partial skipping of the in-frame exon and nuclear accumulation of β-catenin. A single sgRNA can induce small insertions or deletions that partially alter splicing or unexpected larger deletions that remove exons. Exon skipping adds to the unexpected outcomes that must be accounted for, and perhaps taken advantage of, in CRISPR experiments.
CRISPR被广泛用于通过诱导小的插入和缺失来破坏基因功能。在这里，我们表明一些单向导RNA（sgRNA）可以诱导外显子跳跃或删除外显子的大基因组缺失。例如，对编码自抑制结构域的β-连环蛋白外显子3进行CRISPR介导的编辑，会诱导该读码框内的外显子部分跳跃，并导致β-连环蛋白在细胞核中积累。单个sgRNA可以诱导小的插入或缺失，从而部分改变剪接，或者诱导意外的更大缺失，从而删除外显子。外显子跳跃增加了CRISPR实验中必须考虑甚至可能加以利用的意外结果。
