Comprehensive analysis and accurate quantification of unintended large gene modifications induced by CRISPR-Cas9 gene editing

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dc.citation.articleNumbereabo7676
dc.citation.issueNumber42
dc.citation.journalTitleScience Advances
dc.citation.volumeNumber8
dc.contributor.authorPark, So Hyun
dc.contributor.authorCao, Mingming
dc.contributor.authorPan, Yidan
dc.contributor.authorDavis, Timothy H.
dc.contributor.authorSaxena, Lavanya
dc.contributor.authorDeshmukh, Harshavardhan
dc.contributor.authorFu, Yilei
dc.contributor.authorTreangen, Todd
dc.contributor.authorSheehan, Vivien A.
dc.contributor.authorBao, Gang
dc.date.accessioned2022-12-13T19:11:38Z
dc.date.available2022-12-13T19:11:38Z
dc.date.issued2022
dc.description.abstractMost genome editing analyses to date are based on quantifying small insertions and deletions. Here, we show that CRISPR-Cas9 genome editing can induce large gene modifications, such as deletions, insertions, and complex local rearrangements in different primary cells and cell lines. We analyzed large deletion events in hematopoietic stem and progenitor cells (HSPCs) using different methods, including clonal genotyping, droplet digital polymerase chain reaction, single-molecule real-time sequencing with unique molecular identifier, and long-amplicon sequencing assay. Our results show that large deletions of up to several thousand bases occur with high frequencies at the Cas9 on-target cut sites on the HBB (11.7 to 35.4%), HBG (14.3%), and BCL11A (13.2%) genes in HSPCs and the PD-1 (15.2%) gene in T cells. Our findings have important implications to advancing genome editing technologies for treating human diseases, because unintended large gene modifications may persist, thus altering the biological functions and reducing the available therapeutic alleles.
dc.identifier.citationPark, So Hyun, Cao, Mingming, Pan, Yidan, et al.. "Comprehensive analysis and accurate quantification of unintended large gene modifications induced by CRISPR-Cas9 gene editing." <i>Science Advances,</i> 8, no. 42 (2022) AAAS: https://doi.org/10.1126/sciadv.abo7676.
dc.identifier.digitalsciadv-abo7676
dc.identifier.doihttps://doi.org/10.1126/sciadv.abo7676
dc.identifier.urihttps://hdl.handle.net/1911/114129
dc.language.isoeng
dc.publisherAAAS
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.titleComprehensive analysis and accurate quantification of unintended large gene modifications induced by CRISPR-Cas9 gene editing
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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