{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Rananaware SR"],"funding":["University of Florida","NIAID NIH HHS","Exxon Mobil Gator Alumni Faculty","UF Herbert Wertheim College of Engineering","Shah Foundation","NIAID","NIH","NIGMS NIH HHS","NIH HHS"],"pagination":["gkaf887"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC12448901"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["53(17)"],"pubmed_abstract":["CRISPR-Cas12a enzymes are RNA-guided nucleases widely used for programmable genome editing and diagnostics. Perfect complementarity between guide RNA and target DNA is essential for efficient binding and cleavage by Cas12a. However, we report that a particular ortholog of Cas12a, Acidaminococcus sp. Cas12a (AsCas12a), shows an unexpected tolerance to noncomplementary insertions at various positions in its DNA target. AsCas12a remains functional despite DNA bubbles or loops in the CRISPR-RNA (crRNA)-target DNA duplex, displaying both cis- and trans-cleavage activities even when the target harbors insertions of lengths 1-20 nucleotides in the crRNA-binding region. This activity is sequence-independent and works for ssDNA and is observed on dsDNA in vitro for specific insertion lengths/positi"],"journal":["Nucleic acids research"],"pubmed_title":["AsCas12a tolerates insertions in target DNA."],"pmcid":["PMC12448901"],"funding_grant_id":["R21 AI156321","R21 AI168795","R35 GM147788","5R61AI181016","R61 AI181016","R35GM147788","R21AI156321","R21AI168795"],"pubmed_authors":["Karalkar VN","Shoemaker GM","Fang A","Nguyen LT","Vesco EK","Pizzano BLM","Lange IH","Lewis JG","Pedada D","Jain PK","Bodin AP","Sandoval LSW","Antal SG","Rakestraw NR","Rananaware SR","Aguilar D","Meister KS","Flannery SJ"],"additional_accession":[]},"is_claimable":false,"name":"AsCas12a tolerates insertions in target DNA.","description":"CRISPR-Cas12a enzymes are RNA-guided nucleases widely used for programmable genome editing and diagnostics. Perfect complementarity between guide RNA and target DNA is essential for efficient binding and cleavage by Cas12a. However, we report that a particular ortholog of Cas12a, Acidaminococcus sp. Cas12a (AsCas12a), shows an unexpected tolerance to noncomplementary insertions at various positions in its DNA target. AsCas12a remains functional despite DNA bubbles or loops in the CRISPR-RNA (crRNA)-target DNA duplex, displaying both cis- and trans-cleavage activities even when the target harbors insertions of lengths 1-20 nucleotides in the crRNA-binding region. This activity is sequence-independent and works for ssDNA and is observed on dsDNA in vitro for specific insertion lengths/positi","dates":{"release":"2025-01-01T00:00:00Z","publication":"2025 Sep","modification":"2026-06-03T14:39:13.683Z","creation":"2026-05-29T03:05:31.973Z"},"accession":"S-EPMC12448901","cross_references":{"pubmed":["40966510"],"doi":["10.1093/nar/gkaf887"]}}