{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Pandit B"],"funding":["NHGRI NIH HHS","NIGMS NIH HHS"],"pagination":["2551-2556"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC12715751"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["36(12)"],"pubmed_abstract":["CRISPR-Cas12a is a versatile biosensing platform that detects sequence-specific DNA or RNA targets via a CRISPR RNA (crRNA) guide. While Cas12a's specificity is dictated by its crRNA, chemical modifications within the crRNA can influence nuclease performance. Here, we examined the effects of two well-known RNA modifications, N<sup>6</sup>-methyladenosine (m<sup>6</sup>A) and 5-methylcytosine (m<sup>5</sup>C), introduced into the different positions of the guide region of a crRNA. Melting temperature (<i>T</i><sub>m</sub>) analysis showed that m<sup>6</sup>A had a minimal impact on RNA-DNA duplex stability. In contrast, the incorporation of m<sup>5</sup>C residues stabilized the duplex. Using a fluorescence recovery assay, we found that both modifications preserved Cas12a's nuclease activit"],"journal":["Bioconjugate chemistry"],"pubmed_title":["Effects of N&lt;sup&gt;6&lt;/sup&gt;-Methyladenosine (m&lt;sup&gt;6&lt;/sup&gt;A) and 5-Methylcytosine (m&lt;sup&gt;5&lt;/sup&gt;C) Modifications in the Guide Region of CRISPR RNA on Cas12a Nuclease Activity."],"pmcid":["PMC12715751"],"funding_grant_id":["R35 GM156250","R21 HG012257"],"pubmed_authors":["Yigit MV","Royzen M","Pandit B","Yang Q","Hanson E","Dagci H"],"additional_accession":[]},"is_claimable":false,"name":"Effects of N&lt;sup&gt;6&lt;/sup&gt;-Methyladenosine (m&lt;sup&gt;6&lt;/sup&gt;A) and 5-Methylcytosine (m&lt;sup&gt;5&lt;/sup&gt;C) Modifications in the Guide Region of CRISPR RNA on Cas12a Nuclease Activity.","description":"CRISPR-Cas12a is a versatile biosensing platform that detects sequence-specific DNA or RNA targets via a CRISPR RNA (crRNA) guide. While Cas12a's specificity is dictated by its crRNA, chemical modifications within the crRNA can influence nuclease performance. Here, we examined the effects of two well-known RNA modifications, N<sup>6</sup>-methyladenosine (m<sup>6</sup>A) and 5-methylcytosine (m<sup>5</sup>C), introduced into the different positions of the guide region of a crRNA. Melting temperature (<i>T</i><sub>m</sub>) analysis showed that m<sup>6</sup>A had a minimal impact on RNA-DNA duplex stability. In contrast, the incorporation of m<sup>5</sup>C residues stabilized the duplex. Using a fluorescence recovery assay, we found that both modifications preserved Cas12a's nuclease activit","dates":{"release":"2025-01-01T00:00:00Z","publication":"2025 Dec","modification":"2026-06-06T04:48:54.307Z","creation":"2026-05-25T03:12:10.991Z"},"accession":"S-EPMC12715751","cross_references":{"pubmed":["41346237"],"doi":["10.1021/acs.bioconjchem.5c00498"]}}