The CRISPR-Cas9 system can be modified to perform “epigenetic editing” by utilizing the catalytically inactive (dead) Cas9 (dCas9) to recruit regulatory proteins to specific genomic locations. In prior studies, epigenetic editing with multimers of the transactivator VP16 and guide RNAs (gRNAs) was found to cause adverse cellular responses. These side effects may confound studies inducing new cellular properties, especially if the cellular responses are maintained through cell divisions—an epigenetic regulatory property. Here, we show how distinct components of this CRISPR-dCas9 activation system, particularly dCas9 with untargeted gRNAs, upregulate genes associated with transcriptional stress, defense response, and regulation of cell death. Our results highlight a previously undetected acute stress response to CRISPR-dCas9 components in human cells, which is transient and not maintained through cell divisions.

中文翻译：

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CRISPR-dCas9 激活系统诱导的细胞应激反应不能通过细胞分裂遗传


可以修改 CRISPR-Cas9 系统，利用催化失活（死）Cas9 (dCas9) 将调节蛋白招募到特定的基因组位置，从而执行“表观遗传编辑”。在之前的研究中，发现使用反式激活因子 VP16 和引导 RNA (gRNA) 多聚体进行表观遗传编辑会引起不良的细胞反应。这些副作用可能会混淆诱导新细胞特性的研究，特别是如果细胞反应是通过细胞分裂（一种表观遗传调控特性）来维持的。在这里，我们展示了这种 CRISPR-dCas9 激活系统的不同组件，特别是具有非靶向 gRNA 的 dCas9，如何上调与转录应激、防御反应和细胞死亡调节相关的基因。我们的结果强调了人类细胞中先前未检测到的对 CRISPR-dCas9 成分的急性应激反应，这种应激反应是短暂的，并且不会通过细胞分裂维持。