CRISPR-assisted transcription activation by phaseseparation proteins  

作　　者：Jiaqi Liu Yuxi Chen Baoting Nong Xiao Luo Kaixin Cui Zhan Li Pengfei Zhang Wenqiong Tan Yue Yang Wenbin Ma Puping Liang Zhou Songyang 

机构地区：[1]State Key Laboratory of Biocontrol,MOE Key Laboratory of Gene Function and Regulation and Guangzhou Key Laboratory of Healthy Aging Research,School of Life Sciences,Sun Yat-senUniversity,Guangzhou510275,China [2]Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou 510275,China [3]Lumiere Therapeutics,Suzhou215000,China

出　　处：《Protein & Cell》2023年第12期874-887,共14页蛋白质与细胞（英文版）

基　　金：supported by the National Key Research and Development Program of China(2017YFA0102801);the National Natural Science Foundation of China(91640119,81330055,and 32001063);the Guangdong Special Support Program(2019BT02Y276);the Natural Science Foundation of Guangdong Province(2023A1515010176);the Guangzhou Science and Technology Program key projects(2023A04J1952)。

摘　　要：The clustered regularly interspaced short palindromic repeats(CRISPR)-Cas9 system has been widely used for genome engineering and transcriptional regulation in many different organisms.Current CRISPR-activation(CRISPRa)platforms often require multiple components because of inefficient transcriptional activation.Here,we fused different phase-separation proteins to dCas9-VPR(dCas9-VP64-P65-RTA)and observed robust increases in transcriptional activation efficiency.Notably,human NUP98(nucleoporin 98)and FUS(fused in sarcoma)IDR domains were best at enhancing dCas9-VPR activity,with dCas9-VPR-FUS IDR(VPRF)outperforming the other CRISPRa systems tested in this study in both activation efficiency and system simplicity.dCas9-VPRF overcomes the target strand bias and widens gRNA designing windows without affecting the off-target effect of dCas9-VPR.These findings demonstrate the feasibility of using phase-separation proteins to assist in the regulation of gene expression and support the broad appeal of the dCas9-VPRF system in basic and clinical applications.

关 键 词：CRISPR transcriptional activation phase-separation proteins 

分 类 号：Q78[生物学—分子生物学]