Characterization of Argonaute Nuclease from Mesophilic Bacterium <i>Chroococcidiopsis</i>.

ABSTRACT: Mesophilic microbial sources of prokaryotic Argonaute (pAgo) programmable nucleases have garnered considerable attention for their potential applications in genome editing and molecular diagnostics. In this study, we characterized a novel pAgo from the mesophilic bacterium Chroococcidiopsis sp. (ChAgo), which can cleave single-stranded DNA (ssDNA) using both 5'-phosphorylated guide DNA (5'P-gDNA) and 5'-hydroxylated guide DNA (5'OH-gDNA). Efficient cleavage occurs using 14-25 nt 5'P-gDNA and 13-20 nt 5'OH-gDNA in the presence of Mn²⁺ ions at temperatures ranging from 25 to 75 °C, with optimal activity at 55 °C. ChAgo demonstrates low tolerance for single-base mismatches, similar to other pAgo proteins. The cleavage efficiency varies based on the guide/target pair, with mi

SUBMITTER: Peng Y 

PROVIDER: S-EPMC11817465 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature