{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Worle E"],"funding":["Deutsche Forschungsgemeinschaft","Australian-Germany Joint Research co-operation scheme – University Australia/German Academic Exchange Service","Australian Government"],"pagination":["10153-10168"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9508855"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["50(17)"],"pubmed_abstract":["Nucleases derived from the prokaryotic defense system CRISPR-Cas are frequently re-purposed for gene editing and molecular diagnostics. Hence, an in-depth understanding of the molecular mechanisms of these enzymes is of crucial importance. We focused on Cas12a from Francisella novicida (FnCas12a) and investigated the functional role of helix 1, a structural element that together with the bridge helix (BH) connects the recognition and the nuclease lobes of FnCas12a. Helix 1 is structurally connected to the lid domain that opens upon DNA target loading thereby activating the active site of FnCas12a. We probed the structural states of FnCas12a variants altered in helix 1 and/or the bridge helix using single-molecule FRET measurements and assayed the pre-crRNA processing, cis- and trans-DNA cl"],"journal":["Nucleic acids research"],"pubmed_title":["Allosteric activation of CRISPR-Cas12a requires the concerted movement of the bridge helix and helix 1 of the RuvC II domain."],"pmcid":["PMC9508855"],"funding_grant_id":["UA-DAAD 575113331","GR 3840/3-2"],"pubmed_authors":["Grohmann D","Newman A","Worle E","D'Silva J","Burgio G"],"additional_accession":[]},"is_claimable":false,"name":"Allosteric activation of CRISPR-Cas12a requires the concerted movement of the bridge helix and helix 1 of the RuvC II domain.","description":"Nucleases derived from the prokaryotic defense system CRISPR-Cas are frequently re-purposed for gene editing and molecular diagnostics. Hence, an in-depth understanding of the molecular mechanisms of these enzymes is of crucial importance. We focused on Cas12a from Francisella novicida (FnCas12a) and investigated the functional role of helix 1, a structural element that together with the bridge helix (BH) connects the recognition and the nuclease lobes of FnCas12a. Helix 1 is structurally connected to the lid domain that opens upon DNA target loading thereby activating the active site of FnCas12a. We probed the structural states of FnCas12a variants altered in helix 1 and/or the bridge helix using single-molecule FRET measurements and assayed the pre-crRNA processing, cis- and trans-DNA cl","dates":{"release":"2022-01-01T00:00:00Z","publication":"2022 Sep","modification":"2026-05-28T01:09:28.737Z","creation":"2025-04-19T22:49:04.136Z"},"accession":"S-EPMC9508855","cross_references":{"pubmed":["36107767"],"doi":["10.1093/nar/gkac767"]}}