CRISPR-assisted transcription activation by phase-separation proteins.

IF 13.6 1区 生物学 Q1 CELL BIOLOGY
Jiaqi Liu, Yuxi Chen, Baoting Nong, Xiao Luo, Kaixin Cui, Zhan Li, Pengfei Zhang, Wenqiong Tan, Yue Yang, Wenbin Ma, Puping Liang, Zhou Songyang
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引用次数: 2

Abstract

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.

通过相分离蛋白的crispr辅助转录激活。
聚集的规则间隔短回文重复序列(CRISPR)-Cas9系统已广泛用于许多不同生物的基因组工程和转录调控。由于转录激活效率低下,目前的crispr激活(CRISPRa)平台通常需要多个组件。在这里,我们将不同的相分离蛋白与dCas9-VPR (dCas9-VP64-P65-RTA)融合,并观察到转录激活效率的显著提高。值得注意的是,人类NUP98(核孔蛋白98)和FUS(融合在肉瘤中)IDR结构域在增强dCas9-VPR活性方面表现最好,其中dCas9-VPR-FUS IDR (VPRF)在激活效率和系统简单性方面都优于本研究中测试的其他CRISPRa系统。dCas9-VPR克服了靶链偏置,在不影响dCas9-VPR脱靶效应的情况下,拓宽了gRNA的设计窗口。这些发现证明了使用相分离蛋白辅助基因表达调控的可行性,并支持dCas9-VPRF系统在基础和临床应用中的广泛吸引力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Protein & Cell
Protein & Cell CELL BIOLOGY-
CiteScore
24.00
自引率
0.90%
发文量
1029
审稿时长
6-12 weeks
期刊介绍: Protein & Cell is a monthly, peer-reviewed, open-access journal focusing on multidisciplinary aspects of biology and biomedicine, with a primary emphasis on protein and cell research. It publishes original research articles, reviews, and commentaries across various fields including biochemistry, biophysics, cell biology, genetics, immunology, microbiology, molecular biology, neuroscience, oncology, protein science, structural biology, and translational medicine. The journal also features content on research policies, funding trends in China, and serves as a platform for academic exchange among life science researchers.
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