{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Tan R"],"funding":["NIGMS NIH HHS"],"pagination":["852-867.e5"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC8964063"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["82(4)"],"pubmed_abstract":["Leading CRISPR-Cas technologies employ Cas9 and Cas12 enzymes that generate RNA-guided dsDNA breaks. Yet, the most abundant microbial adaptive immune systems, Type I CRISPRs, are under-exploited for eukaryotic applications. Here, we report the adoption of a minimal CRISPR-Cas3 from Neisseria lactamica (Nla) type I-C system to create targeted large deletions in the human genome. RNP delivery of its processive Cas3 nuclease and target recognition complex Cascade can confer ∼95% editing efficiency. Unexpectedly, NlaCascade assembly in bacteria requires internal translation of a hidden component Cas11 from within the cas8 gene. Furthermore, expressing a separately encoded NlaCas11 is the key to enable plasmid- and mRNA-based editing in human cells. Finally, we demonstrate that supplying cas11 is a universal strategy to systematically implement divergent I-C, I-D, and I-B CRISPR-Cas3 editors with compact sizes, distinct PAM preferences, and guide orthogonality. These findings greatly expand our ability to engineer long-range genome edits."],"journal":["Molecular cell"],"pubmed_title":["Cas11 enables genome engineering in human cells with compact CRISPR-Cas3 systems."],"pmcid":["PMC8964063"],"funding_grant_id":["R35 GM137883","K99 GM117268","R00 GM117268","R35 GM118174"],"pubmed_authors":["Hou Z","Gramelspacher MJ","Zhang Y","Xiao Y","Ke A","Zhou X","Tan R","Krueger RK"],"additional_accession":[]},"is_claimable":false,"name":"Cas11 enables genome engineering in human cells with compact CRISPR-Cas3 systems.","description":"Leading CRISPR-Cas technologies employ Cas9 and Cas12 enzymes that generate RNA-guided dsDNA breaks. Yet, the most abundant microbial adaptive immune systems, Type I CRISPRs, are under-exploited for eukaryotic applications. Here, we report the adoption of a minimal CRISPR-Cas3 from Neisseria lactamica (Nla) type I-C system to create targeted large deletions in the human genome. RNP delivery of its processive Cas3 nuclease and target recognition complex Cascade can confer ∼95% editing efficiency. Unexpectedly, NlaCascade assembly in bacteria requires internal translation of a hidden component Cas11 from within the cas8 gene. Furthermore, expressing a separately encoded NlaCas11 is the key to enable plasmid- and mRNA-based editing in human cells. Finally, we demonstrate that supplying cas11 is a universal strategy to systematically implement divergent I-C, I-D, and I-B CRISPR-Cas3 editors with compact sizes, distinct PAM preferences, and guide orthogonality. These findings greatly expand our ability to engineer long-range genome edits.","dates":{"release":"2022-01-01T00:00:00Z","publication":"2022 Feb","modification":"2025-04-20T03:04:12.735Z","creation":"2025-04-20T03:04:12.735Z"},"accession":"S-EPMC8964063","cross_references":{"pubmed":["35051351"],"doi":["10.1016/j.molcel.2021.12.032"]}}