Massively parallel characterization of CRISPR activator efficacy in human induced pluripotent stem cells and neurons

文献类型： 外文期刊

第一作者： Wu, Qianxin

作者： Wu, Qianxin;Wu, Junjing;Chen, Xi;Iwama, Sho;Carobbio, Stefania;Keen, Peter;Vidal-Puig, Antonio;Bassett, Andrew;Karim, Kaiser;Kotter, Mark R.;Wu, Junjing;Chen, Xi;Wang, Tengyao;Carobbio, Stefania;Vidal-Puig, Antonio;Carobbio, Stefania;Vidal-Puig, Antonio

作者机构：

期刊名称：MOLECULAR CELL （ 影响因子：16.0； 五年影响因子：18.4 ）

ISSN： 1097-2765

年卷期： 2023 年 83 卷 7 期

页码：

收录情况： SCI

摘要： CRISPR activation (CRISPRa) is an important tool to perturb transcription, but its effectiveness varies be-tween target genes. We employ human pluripotent stem cells with thousands of randomly integrated bar-coded reporters to assess epigenetic features that influence CRISPRa efficacy. Basal expression levels are influenced by genomic context and dramatically change during differentiation to neurons. Gene activa-tion by dCas9-VPR is successful in most genomic contexts, including developmentally repressed regions, and activation level is anti-correlated with basal gene expression, whereas dCas9-p300 is ineffective in stem cells. Certain chromatin states, such as bivalent chromatin, are particularly sensitive to dCas9-VPR, whereas constitutive heterochromatin is less responsive. We validate these rules at endogenous genes and show that activation of certain genes elicits a change in the stem cell transcriptome, sometimes showing features of differentiated cells. Our data provide rules to predict CRISPRa outcome and highlight its utility to screen for factors driving stem cell differentiation.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序