The prokaryotic clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated (CRISPR/Cas) protein system has been reconstructed for a revolutionary targeted genome modification platform and nucleic acid detection tool. Restricted by the protospacer adjacent motif (PAM) requirement, specificity, and efficiency, more Cas proteins need to be characterized and adapted for genome editing and other applications. The RNA-guided DNA interference activity and ssDNA (single-strand DNA) trans-cleavage activity of Cas12c (subtypes V–C) has not been reconstructed in vitro, limiting its genome editing and nucleic acid detection applications.