CRISPR-Cas12a has both cis-and trans-cleavage activities on single-stranded DNA
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Dear Editor, The CRISPR-associated protein Cas12a (previously known as Cpf1), which is an endonuclease from the type V-A CRISPR system, has been applied in both in vivo genome editing and in vitro DNA assembly.1-3 Cas12a is guided by a single CRISPR RNA (crRNA) with a T-rich protospacer adjacent motif (PAM) sequence to cleave double-stranded DNA (dsDNA) targets, generating sticky ends. Different from Cas9, Cas12a cleaves both the target and non-target strands of a targeted dsDNA by a single active site in the RuvC catalytic pocket4-6 (Supplementary information, Figure S12a). Besides, Cas12a also processes precursor crRNAs to generate mature crRNAs.7 However, the cleavage activity of Cas12a on single-stranded DNA (ssDNA) targets is less understood.
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We thank Chao Lei and Qi Li for their helpful discussion. This work was supported by grants from the Strategic Priority Research Program of the Chinese Academy of SciencesXDB19040200;the Youth Innovation Promotion Association CAS2017322;the National Natural Science Foundation of China31421061,31430004 and 31300031