2022
Structural Basis for Reduced Dynamics of Three Engineered HNH Endonuclease Lys-to-Ala Mutants for the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-Associated 9 (CRISPR/Cas9) Enzyme
Wang J, Skeens E, Arantes PR, Maschietto F, Allen B, Kyro GW, Lisi GP, Palermo G, Batista VS. Structural Basis for Reduced Dynamics of Three Engineered HNH Endonuclease Lys-to-Ala Mutants for the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-Associated 9 (CRISPR/Cas9) Enzyme. Biochemistry 2022, 61: 785-794. PMID: 35420793, PMCID: PMC9069930, DOI: 10.1021/acs.biochem.2c00127.Peer-Reviewed Original ResearchMeSH KeywordsClustered Regularly Interspaced Short Palindromic RepeatsCRISPR-Associated Protein 9CRISPR-Cas SystemsDNAEndonucleasesConceptsShort palindromic repeatsSubstrate specificityPalindromic repeatsAla mutantWT enzymeRNA-binding domainAssociated 9 (Cas9) systemForeign DNA sequencesDNA strandsWild-type enzymeDouble-strand breaksEnhanced substrate specificityHNH active siteDynamics of proteinsType II immunityCas9 proteinDNA substratesDNA sequencesStructural basisMutantsAla substitutionDistinct conformationsSingle LysCatalytic siteEnzyme
2021
Enhanced specificity mutations perturb allosteric signaling in CRISPR-Cas9
Nierzwicki L, East K, Morzan U, Arantes P, Batista V, Lisi G, Palermo G. Enhanced specificity mutations perturb allosteric signaling in CRISPR-Cas9. ELife 2021, 10: e73601. PMID: 34908530, PMCID: PMC8741213, DOI: 10.7554/elife.73601.Peer-Reviewed Original ResearchMeSH KeywordsAllosteric RegulationCRISPR-Cas SystemsGenetic VariationGenotypeMolecular Dynamics SimulationMolecular StructureMutationStreptococcus pyogenesConceptsHNH domainAllosteric communicationCatalytic HNH domainDNA recognition regionSpecificity-enhancing mutationsGenome editing capabilitiesAllosteric signalingAllosteric signalMutations perturbAllosteric hotspotsSpecificity enhancementCas9 endonucleaseMutational studiesDNA recognitionAllosteric connectivityAllosteric roleMolecular toolsAllosteric structureRecognition regionMolecular levelBiochemical studiesDNA cleavageSolution NMRMutationsCatalytic site