上海科技大学知识管理系统

Cas12f possesses both cis- and trans-cleavage activities, with the former being extensively studied for its application in genome edit-ing, while the latter remains less explored, particularly for diagnostic purposes, and is mostly focused on Un1Cas12f1. In this study, weconducted a comprehensive comparison of the trans-cleavage activities of four characterized Cas12f proteins, demonstrating that allfour exhibit trans-DNase activity triggered by double-stranded DNA (dsDNA), single-stranded DNA (ssDNA), and single-stranded RNA(ssRNA). Additionally, we identified distinct base preferences for trans-cleavage substrates among these proteins. Our further investi-gation into the activities of Cas12f revealed the intricate relationship between cis- and trans-cleavage activities under various conditions. Our study provides a multifaceted characterization of the trans-cleavage features of Cas12f nucleases, offering new avenues fora deeper comprehension of the mechanisms underlying Cas12f's functionality.