{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Marina RJ"],"funding":["NEI NIH HHS","NIH/NINDS Ruth L. Kirschstein National Research Service Award","NHGRI NIH HHS","NINDS NIH HHS","NIH SIG grant","NIH/NINDS Career Transition Award","NIH","National Institute of General Medical Sciences","NIGMS NIH HHS","NIH HHS"],"pagination":["108350"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC8985550"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["33(5)"],"pubmed_abstract":["Site-directed RNA editing approaches offer great potential to correct genetic mutations in somatic cells while avoiding permanent off-target genomic edits. Nuclease-dead RNA-targeting CRISPR-Cas systems recruit functional effectors to RNA molecules in a programmable fashion. Here, we demonstrate a Streptococcus pyogenes Cas9-ADAR2 fusion system that uses a 3' modified guide RNA (gRNA) to enable adenosine-to-inosine (A-to-I) editing of specific bases on reporter and endogenously expressed mRNAs. Due to the sufficient nature of the 3' gRNA extension sequence, we observe that Cas9 gRNA spacer sequences are dispensable for directed RNA editing, revealing that Cas9 can act as an RNA-aptamer-binding protein. We demonstrate that Cas9-based A-to-I editing is comparable in on-target efficiency and "],"journal":["Cell reports"],"pubmed_title":["Evaluation of Engineered CRISPR-Cas-Mediated Systems for Site-Specific RNA Editing."],"pmcid":["PMC8985550"],"funding_grant_id":["S10OD026929","R01NS103172","K22NS112678","T32 GM008666","R01 HG004659","F31 NS111859","R01HG004659","T32GM008666","R01 NS103172","S10 OD026929","K22 NS112678","T32 GM145427","R01EY029166","F31NS111859","R01 EY029166"],"pubmed_authors":["Brannan KW","Dong KD","Yee BA","Yeo GW","Marina RJ"],"additional_accession":[]},"is_claimable":false,"name":"Evaluation of Engineered CRISPR-Cas-Mediated Systems for Site-Specific RNA Editing.","description":"Site-directed RNA editing approaches offer great potential to correct genetic mutations in somatic cells while avoiding permanent off-target genomic edits. Nuclease-dead RNA-targeting CRISPR-Cas systems recruit functional effectors to RNA molecules in a programmable fashion. Here, we demonstrate a Streptococcus pyogenes Cas9-ADAR2 fusion system that uses a 3' modified guide RNA (gRNA) to enable adenosine-to-inosine (A-to-I) editing of specific bases on reporter and endogenously expressed mRNAs. Due to the sufficient nature of the 3' gRNA extension sequence, we observe that Cas9 gRNA spacer sequences are dispensable for directed RNA editing, revealing that Cas9 can act as an RNA-aptamer-binding protein. We demonstrate that Cas9-based A-to-I editing is comparable in on-target efficiency and ","dates":{"release":"2020-01-01T00:00:00Z","publication":"2020 Nov","modification":"2026-05-30T15:04:38.451Z","creation":"2025-04-04T23:04:15.233Z"},"accession":"S-EPMC8985550","cross_references":{"pubmed":["33147453"],"doi":["10.1016/j.celrep.2020.108350"]}}