Comparison of DNA targeting CRISPR editors in human cells

Cell & Bioscience

Research

Linxuan Huang¹  Zhihong Tan²  Guanjie Huang²  Xin Zhang³  Shufeng Ma⁴  Hongxin Huang⁵  Ying Lin⁶  Jinhe Li⁷  Weiqi Lv⁸  Yongfei Hu⁸ 

[1] Affiliated Dongguan Hospital, Southern Medical University, (Dongguan People’s Hospital), 523058, Dongguan, China;Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, 510515, Guangzhou, China;Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, 510515, Guangzhou, China;Affiliated Dongguan Hospital, Southern Medical University, (Dongguan People’s Hospital), 523058, Dongguan, China;Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, 510515, Guangzhou, China;Department of Nephrology, Shenzhen Hospital, Southern Medical University, 518110, Shenzhen, China;Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, 510515, Guangzhou, China;Dermatology Hospital, Southern Medical University, 510091, Guangzhou, China;Affiliated Cancer Hospital & Institute of Guangzhou Medical University, 510095, Guangzhou, China;Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, 510515, Guangzhou, China;Experimental Education/Administration Center, School of Basic Medical Science, Southern Medical University, 510515, Guangzhou, China;Department of Anesthesiology, The Third Affiliated Hospital of Southern Medical University, 510630, Guangzhou, China;Dermatology Hospital, Southern Medical University, 510091, Guangzhou, China;

关键词: Cas12f1 nuclease;    CRISPR-Cas system;    DNA targeting;    Specificity;    Comparison;   

DOI  :  10.1186/s13578-023-00958-z

 received in 2022-10-21, accepted in 2023-01-05,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundProfiling and comparing the performance of current widely used DNA targeting CRISPR systems provide the basic information for the gene-editing toolkit and can be a useful resource for this field. In the current study, we made a parallel comparison between the recently reported miniature Cas12f1 (Un1Cas12f1 and AsCas12f1) and the widely used Cas12a and Cas9 nucleases in mammalian cells.ResultsWe found that as a CRISPRa activator, Un1Cas12f1 could induce gene expression with a comparable level to that of Cas12a and Cas9, while as a DNA cleavage editor, Cas12f1 exhibited similar properties to Cas12a, like high specificity and dominantly induced deletions over insertions, but with less activity. In contrast, wild-type SpCas9 showed the highest activity, lowest specificity, and induced balanced deletions and insertions. Thus, Cas12f1 is recommended for gene-activation-based applications, Cas12a is for therapy applications, and wild-type Cas9 is for in vitro and animal investigations.ConclusionThe comparison provided the editing properties of the widely used DNA-targeting CRISPR systems in the gene-editing field.

【 授权许可】

CC BY   
© The Author(s) 2023

【 图 表 】

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