Nature Chemical Biology. 2021 Nov
Zhaowei Wu, Yifei Zhang , Haopeng Yu, Deng Pan, Yujue Wang, Yannan Wang, Fan Li, Chang Liu, Hao Nan, Weizhong Chen, Quanjiang Ji
Products used in the paper Details Operation
AAV vector packaging The AAV production was carried out by PackGene Biotech Co., giving a final physical titration of 1013 GC ml−1 (GC, genome copy number). Request Quote

Research Field: genome editing

AAV Serotype: AAV2


The RNA-guided CRISPR-associated (Cas) nucleases are versatile tools for genome editing in various organisms. The large sizes of the commonly used Cas9 and Cas12a nucleases restrict their flexibility in therapeutic applications that use the cargo-size-limited adeno-associated virus delivery vehicle. More compact systems would thus offer more therapeutic options and functionality for this field. Here, we report a miniature class 2 type V-F CRISPR-Cas genome-editing system from Acidibacillus sulfuroxidans (AsCas12f1, 422 amino acids). AsCas12f1 is an RNA-guided endonuclease that recognizes 5′ T-rich protospacer adjacent motifs and creates staggered double-stranded breaks to target DNA. We show that AsCas12f1 functions as an effective genome-editing tool in both bacteria and human cells using various delivery methods, including plasmid, ribonucleoprotein and adeno-associated virus. The small size of AsCas12f1 offers advantages for cellular delivery, and characterizations of AsCas12f1 may facilitate engineering more compact genome-manipulation technologies.

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