{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Liang SQ"],"funding":["American Cancer Society","NCATS NIH HHS","NIA NIH HHS","NHLBI NIH HHS","NHGRI NIH HHS","NCI NIH HHS","NIGMS NIH HHS"],"pagination":["437"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC8782884"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["13(1)"],"pubmed_abstract":["Analysis of off-target editing is an important aspect of the development of safe nuclease-based genome editing therapeutics. in vivo assessment of nuclease off-target activity has primarily been indirect (based on discovery in vitro, in cells or via computational prediction) or through ChIP-based detection of double-strand break (DSB) DNA repair factors, which can be cumbersome. Herein we describe GUIDE-tag, which enables one-step, off-target genome editing analysis in mouse liver and lung. The GUIDE-tag system utilizes tethering between the Cas9 nuclease and the DNA donor to increase the capture rate of nuclease-mediated DSBs and UMI incorporation via Tn5 tagmentation to avoid PCR bias. These components can be delivered as SpyCas9-mSA ribonucleoprotein complexes and biotin-dsDNA donor for in vivo editing analysis. GUIDE-tag enables detection of off-target sites where editing rates are ≥ 0.2%. UDiTaS analysis utilizing the same tagmented genomic DNA detects low frequency translocation events with off-target sites and large deletions in vivo. The SpyCas9-mSA and biotin-dsDNA system provides a method to capture DSB loci in vivo in a variety of tissues with a workflow that is amenable to analysis of gross genomic alterations that are associated with genome editing."],"journal":["Nature communications"],"pubmed_title":["Genome-wide detection of CRISPR editing in vivo using GUIDE-tag."],"pmcid":["PMC8782884"],"funding_grant_id":["P01 HL131471","R01 AG040061","T32 GM107000","129056-RSG-16-093","R01 GM115911","F30 CA239483","UG3 HL147367","DP2 HL137167","T32 CA130807","R01 HL150669","UG3 TR002668","U24 HG010423"],"pubmed_authors":["Dong X","Liu P","Lee J","Liang SQ","Wolfe SA","Xue W","Haynes CM","Maitland S","Zhu LJ","Watts JK","Sontheimer EJ","Smith JL","Yang Q","Mintzer E"],"additional_accession":[]},"is_claimable":false,"name":"Genome-wide detection of CRISPR editing in vivo using GUIDE-tag.","description":"Analysis of off-target editing is an important aspect of the development of safe nuclease-based genome editing therapeutics. in vivo assessment of nuclease off-target activity has primarily been indirect (based on discovery in vitro, in cells or via computational prediction) or through ChIP-based detection of double-strand break (DSB) DNA repair factors, which can be cumbersome. Herein we describe GUIDE-tag, which enables one-step, off-target genome editing analysis in mouse liver and lung. The GUIDE-tag system utilizes tethering between the Cas9 nuclease and the DNA donor to increase the capture rate of nuclease-mediated DSBs and UMI incorporation via Tn5 tagmentation to avoid PCR bias. These components can be delivered as SpyCas9-mSA ribonucleoprotein complexes and biotin-dsDNA donor for in vivo editing analysis. GUIDE-tag enables detection of off-target sites where editing rates are ≥ 0.2%. UDiTaS analysis utilizing the same tagmented genomic DNA detects low frequency translocation events with off-target sites and large deletions in vivo. The SpyCas9-mSA and biotin-dsDNA system provides a method to capture DSB loci in vivo in a variety of tissues with a workflow that is amenable to analysis of gross genomic alterations that are associated with genome editing.","dates":{"release":"2022-01-01T00:00:00Z","publication":"2022 Jan","modification":"2025-04-04T19:53:50.755Z","creation":"2025-04-04T19:53:50.755Z"},"accession":"S-EPMC8782884","cross_references":{"pubmed":["35064134"],"doi":["10.1038/s41467-022-28135-9"]}}