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新挑战:Cas9可能增加促癌风险
作者:小柯机器人 发布时间:2020/5/19 13:28:21

美国博德研究所Uri Ben-David小组发现Cas9能够激活p53通路并选择p53失活突变。相关论文于2020年5月18日在线发表在《自然—遗传学》杂志上。

Cas9通常被引入到细胞系中来实现CRISPR–Cas9介导的基因组编辑。在本项研究中,研究人员探索了Cas9本身表达所产生的遗传和转录后果。165组人类癌细胞系以及相应表达Cas9的衍生细胞系的基因表达谱揭示了引入Cas9后p53通路的上调,特别是在野生型TP53(TP53-WT)细胞系中。这在信使RNA和蛋白质水平得到证实。
 
此外,在表达Cas9的细胞系中观察到DNA修复水平升高。42组细胞系的遗传特征表明,Cas9的引入可以导致p53失活突变的出现和扩增。这在同基因TP53-WT和TP53-null细胞系中的竞争实验中得到证实。
 
最后,Cas9在TP53-WT中的活性低于在TP53突变细胞系中的活性,并且Cas9诱导的p53途径激活能够影响细胞对遗传和化学干扰的敏感性。这些发现可能对正确使用CRISPR–Cas9介导的基因组编辑具有广泛的意义。
 
附:英文原文

Title: Cas9 activates the p53 pathway and selects for p53-inactivating mutations

Author: Oana M. Enache, Veronica Rendo, Mai Abdusamad, Daniel Lam, Desiree Davison, Sangita Pal, Naomi Currimjee, Julian Hess, Sasha Pantel, Anwesha Nag, Aaron R. Thorner, John G. Doench, Francisca Vazquez, Rameen Beroukhim, Todd R. Golub, Uri Ben-David

Issue&Volume: 2020-05-18

Abstract: Cas9 is commonly introduced into cell lines to enable CRISPR–Cas9-mediated genome editing. Here, we studied the genetic and transcriptional consequences of Cas9 expression itself. Gene expression profiling of 165 pairs of human cancer cell lines and their Cas9-expressing derivatives revealed upregulation of the p53 pathway upon introduction of Cas9, specifically in wild-type TP53 (TP53-WT) cell lines. This was confirmed at the messenger RNA and protein levels. Moreover, elevated levels of DNA repair were observed in Cas9-expressing cell lines. Genetic characterization of 42 cell line pairs showed that introduction of Cas9 can lead to the emergence and expansion of p53-inactivating mutations. This was confirmed by competition experiments in isogenic TP53-WT and TP53-null (TP53/) cell lines. Lastly, Cas9 was less active in TP53-WT than in TP53-mutant cell lines, and Cas9-induced p53 pathway activation affected cellular sensitivity to both genetic and chemical perturbations. These findings may have broad implications for the proper use of CRISPR–Cas9-mediated genome editing.

DOI: 10.1038/s41588-020-0623-4

Source: https://www.nature.com/articles/s41588-020-0623-4

期刊信息

Nature Genetics:《自然—遗传学》,创刊于1992年。隶属于施普林格·自然出版集团,最新IF:25.455
官方网址:https://www.nature.com/ng/
投稿链接:https://mts-ng.nature.com/cgi-bin/main.plex