{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["35(2)"],"submitter":["Weber JA"],"pubmed_abstract":["Clinical application of CRISPR-Cas9 technology for large deletions of somatic mutations is inefficient, and methods to improve utility suffer from our inability to rapidly assess mono- vs. biallelic deletions. Here we establish a model system for investigating allelic heterogeneity at the single-cell level and identify indel scarring from non-simultaneous nuclease activity at gRNA cut sites as a major barrier to CRISPR-del efficacy both <i>in vitro</i> and <i>in vivo</i>. We show that non-simultaneous nuclease activity is partially prevented via restriction of CRISPR-Cas9 expression via inducible adeno-associated viruses (AAVs) or lipid nanoparticles (LNPs). Inducible AAV-based expression of CRISPR-del machinery significantly improved mono- and biallelic deletion frequency <i>in vivo</i>, "],"journal":["Molecular therapy. Nucleic acids"],"pagination":["102172"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11229411"],"repository":["biostudies-literature"],"pubmed_title":["Temporal restriction of Cas9 expression improves CRISPR-mediated deletion efficacy and fidelity."],"pmcid":["PMC11229411"],"pubmed_authors":["Carrell EM","Lang JF","Davidson BL","Alameh MG","Weber JA"],"additional_accession":[]},"is_claimable":false,"name":"Temporal restriction of Cas9 expression improves CRISPR-mediated deletion efficacy and fidelity.","description":"Clinical application of CRISPR-Cas9 technology for large deletions of somatic mutations is inefficient, and methods to improve utility suffer from our inability to rapidly assess mono- vs. biallelic deletions. Here we establish a model system for investigating allelic heterogeneity at the single-cell level and identify indel scarring from non-simultaneous nuclease activity at gRNA cut sites as a major barrier to CRISPR-del efficacy both <i>in vitro</i> and <i>in vivo</i>. We show that non-simultaneous nuclease activity is partially prevented via restriction of CRISPR-Cas9 expression via inducible adeno-associated viruses (AAVs) or lipid nanoparticles (LNPs). Inducible AAV-based expression of CRISPR-del machinery significantly improved mono- and biallelic deletion frequency <i>in vivo</i>, ","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 Jun","modification":"2025-04-04T11:24:19.75Z","creation":"2025-04-04T11:24:19.75Z"},"accession":"S-EPMC11229411","cross_references":{"pubmed":["38978694"],"doi":["10.1016/j.omtn.2024.102172"]}}