{"date_updated":"2024-06-04T06:55:16Z","publication":"bioRxiv","abstract":[{"lang":"eng","text":"Cas12a2 is a CRISPR-associated nuclease that performs RNA-guided degradation of non-specific single-stranded (ss)RNA, ssDNA and double-stranded (ds)DNA upon recognition of a complementary RNA target, culminating in abortive infection (Dmytrenko 2022). Here, we report structures of Cas12a2 in binary, ternary, and quaternary complexes to reveal a complete activation pathway. Our structures reveal that Cas12a2 is autoinhibited until binding a cognate RNA target, which exposes the RuvC active site within a large, positively charged cleft. Double-stranded DNA substrates are captured through duplex distortion and local melting, stabilized by pairs of ‘aromatic clamp’ residues that are crucial for dsDNA degradation and in <jats:italic>vivo</jats:italic> immune system function. Our work provides a structural basis for this unprecedented mechanism of abortive infection to achieve population-level immunity, which can be leveraged to create rational mutants that degrade a spectrum of collateral substrates."}],"status":"public","date_created":"2024-06-04T06:44:59Z","day":"13","year":"2022","date_published":"2022-06-13T00:00:00Z","article_processing_charge":"No","language":[{"iso":"eng"}],"author":[{"last_name":"Bravo","id":"96aecfa5-8931-11ee-af30-aa6a5d6eee0e","orcid":"0000-0003-0456-0753","first_name":"Jack Peter Kelly","full_name":"Bravo, Jack Peter Kelly"},{"first_name":"Thom","full_name":"Hallmark, Thom","last_name":"Hallmark"},{"last_name":"Naegle","full_name":"Naegle, Bronson","first_name":"Bronson"},{"last_name":"Beisel","full_name":"Beisel, Chase L.","first_name":"Chase L."},{"first_name":"Ryan N.","full_name":"Jackson, Ryan N.","last_name":"Jackson"},{"last_name":"Taylor","first_name":"David W.","full_name":"Taylor, David W."}],"extern":"1","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","main_file_link":[{"open_access":"1","url":"https://doi.org/10.1101/2022.06.13.495754"}],"oa":1,"publisher":"Cold Spring Harbor Laboratory","publication_status":"published","doi":"10.1101/2022.06.13.495754","_id":"17117","month":"06","title":"Large-scale structural rearrangements unleash indiscriminate nuclease activity of CRISPR-Cas12a2","oa_version":"Preprint","citation":{"mla":"Bravo, Jack Peter Kelly, et al. “Large-Scale Structural Rearrangements Unleash Indiscriminate Nuclease Activity of CRISPR-Cas12a2.” <i>BioRxiv</i>, Cold Spring Harbor Laboratory, 2022, doi:<a href=\"https://doi.org/10.1101/2022.06.13.495754\">10.1101/2022.06.13.495754</a>.","ieee":"J. P. K. Bravo, T. Hallmark, B. Naegle, C. L. Beisel, R. N. Jackson, and D. W. Taylor, “Large-scale structural rearrangements unleash indiscriminate nuclease activity of CRISPR-Cas12a2,” <i>bioRxiv</i>. Cold Spring Harbor Laboratory, 2022.","chicago":"Bravo, Jack Peter Kelly, Thom Hallmark, Bronson Naegle, Chase L. Beisel, Ryan N. Jackson, and David W. Taylor. “Large-Scale Structural Rearrangements Unleash Indiscriminate Nuclease Activity of CRISPR-Cas12a2.” <i>BioRxiv</i>. Cold Spring Harbor Laboratory, 2022. <a href=\"https://doi.org/10.1101/2022.06.13.495754\">https://doi.org/10.1101/2022.06.13.495754</a>.","ista":"Bravo JPK, Hallmark T, Naegle B, Beisel CL, Jackson RN, Taylor DW. 2022. Large-scale structural rearrangements unleash indiscriminate nuclease activity of CRISPR-Cas12a2. bioRxiv, <a href=\"https://doi.org/10.1101/2022.06.13.495754\">10.1101/2022.06.13.495754</a>.","ama":"Bravo JPK, Hallmark T, Naegle B, Beisel CL, Jackson RN, Taylor DW. Large-scale structural rearrangements unleash indiscriminate nuclease activity of CRISPR-Cas12a2. <i>bioRxiv</i>. 2022. doi:<a href=\"https://doi.org/10.1101/2022.06.13.495754\">10.1101/2022.06.13.495754</a>","apa":"Bravo, J. P. K., Hallmark, T., Naegle, B., Beisel, C. L., Jackson, R. N., &#38; Taylor, D. W. (2022). Large-scale structural rearrangements unleash indiscriminate nuclease activity of CRISPR-Cas12a2. <i>bioRxiv</i>. Cold Spring Harbor Laboratory. <a href=\"https://doi.org/10.1101/2022.06.13.495754\">https://doi.org/10.1101/2022.06.13.495754</a>","short":"J.P.K. Bravo, T. Hallmark, B. Naegle, C.L. Beisel, R.N. Jackson, D.W. Taylor, BioRxiv (2022)."},"type":"preprint"}