Repurposing type I–F CRISPR–Cas system as a transcriptional activation tool in human cells

Yuxi Chen, Jiaqi Liu, Shengyao Zhi, Qi Zheng, Wenbin Ma, Junjiu Huang, Yizhi Liu, Dan Liu, Puping Liang () and Zhou Songyang ()
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Yuxi Chen: Sun Yat-sen University; MOE Key Laboratory of Gene Function and Regulation and Guangzhou Key Laboratory of Healthy Aging Research, School of Life Sciences, Sun Yat-sen University
Jiaqi Liu: Sun Yat-sen University; MOE Key Laboratory of Gene Function and Regulation and Guangzhou Key Laboratory of Healthy Aging Research, School of Life Sciences, Sun Yat-sen University
Shengyao Zhi: Sun Yat-sen University; MOE Key Laboratory of Gene Function and Regulation and Guangzhou Key Laboratory of Healthy Aging Research, School of Life Sciences, Sun Yat-sen University
Qi Zheng: Sun Yat-sen University; MOE Key Laboratory of Gene Function and Regulation and Guangzhou Key Laboratory of Healthy Aging Research, School of Life Sciences, Sun Yat-sen University
Wenbin Ma: Sun Yat-sen University; MOE Key Laboratory of Gene Function and Regulation and Guangzhou Key Laboratory of Healthy Aging Research, School of Life Sciences, Sun Yat-sen University
Junjiu Huang: Sun Yat-sen University; MOE Key Laboratory of Gene Function and Regulation and Guangzhou Key Laboratory of Healthy Aging Research, School of Life Sciences, Sun Yat-sen University
Yizhi Liu: Sun Yat-sen University
Dan Liu: Baylor College of Medicine, One Baylor Plaza
Puping Liang: Sun Yat-sen University; MOE Key Laboratory of Gene Function and Regulation and Guangzhou Key Laboratory of Healthy Aging Research, School of Life Sciences, Sun Yat-sen University
Zhou Songyang: Sun Yat-sen University; MOE Key Laboratory of Gene Function and Regulation and Guangzhou Key Laboratory of Healthy Aging Research, School of Life Sciences, Sun Yat-sen University

Nature Communications, 2020, vol. 11, issue 1, 1-14

Abstract: Abstract Class 2 CRISPR–Cas proteins have been widely developed as genome editing and transcriptional regulating tools. Class 1 type I CRISPR–Cas constitutes ~60% of all the CRISPR–Cas systems. However, only type I–B and I–E systems have been used to control mammalian gene expression and for genome editing. Here we demonstrate the feasibility of using type I–F system to regulate human gene expression. By fusing transcription activation domain to Pseudomonas aeruginosa type I–F Cas proteins, we activate gene transcription in human cells. In most cases, type I–F system is more efficient than other CRISPR-based systems. Transcription activation is enhanced by elongating the crRNA. In addition, we achieve multiplexed gene activation with a crRNA array. Furthermore, type I–F system activates target genes specifically without off-target transcription activation. These data demonstrate the robustness and programmability of type I–F CRISPR–Cas in human cells.

Date: 2020
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DOI: 10.1038/s41467-020-16880-8

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