Discovery of the widespread site-specific single-stranded nuclease family Ssn

• DOI: 10.1038/s41467-025-57514-1
• Publication Date: 2025-03-10T19:29:56Z
• Authors: Martin Chenal; Alex Rivera-Millot; Luke B. Harrison; Ahmed S. Khairalla; Cecilia Nieves; Ève Bernet; Mansoore Esmaili; Manel Belkhir; Jonathan Perreault; Frédéric J. Veyrier
• Keywords: Nuclease

Site-specific endonucleases that exclusively cut single-stranded DNA have hitherto never been described and constitute a barrier to the development of ssDNA-based technologies. We identify characterize one such family, from GIY-YIG superfamily, widely distributed site-specific nucleases (Ssn) exhibiting unique ssDNA cleavage properties. By first comprehensively studying Ssn homolog Neisseria meningitidis, we demonstrate it interacts specifically with sequence (called NTS) present in hundreds copies surrounding important genes pathogenic Neisseria. In this species, NTS/Ssn interactions modulate natural transformation thus an additional mechanism shaping genome dynamics. further thousands homologs demonstrate, vitro, range nuclease specificities for their corresponding sequence. proofs concept applications including detection digestion RCA. This discovery its set stage innovative molecular tools Here authors family enzymes could be basis tools.