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Detection of unintended on-target effects in CRISPR genome editing by DNA donors carrying diagnostic substitutions
https://oist.repo.nii.ac.jp/records/2918
https://oist.repo.nii.ac.jp/records/2918b4587e16-e564-4686-932a-585026b4454e
名前 / ファイル | ライセンス | アクション |
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Lackner-Detection of unintended on-target effe (2.5 MB)
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Creative Commons Attribution-NonCommercial 4.0 International(https://creativecommons.org/licenses/by-nc/4.0/)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2023-04-20 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | Detection of unintended on-target effects in CRISPR genome editing by DNA donors carrying diagnostic substitutions | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者(英) |
Lackner, Martin
× Lackner, Martin× Helmbrecht, Nelly× Pääbo, Svante× Riesenberg, Stephan |
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書誌情報 |
en : Nucleic Acids Research 巻 51, 号 5, p. e26, 発行日 2023-01-09 |
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抄録 | ||||||
内容記述タイプ | Other | |||||
内容記述 | CRISPR nucleases can introduce double-stranded DNA breaks in genomes at positions specified by guide RNAs. When repaired by the cell, this may result in the introduction of insertions and deletions or nucleotide substitutions provided by exogenous DNA donors. However, cellular repair can also result in unintended on-target effects, primarily larger deletions and loss of heterozygosity due to gene conversion. Here we present a strategy that allows easy and reliable detection of unintended on-target effects as well as the generation of control cells that carry wild-type alleles but have demonstratively undergone genome editing at the target site. Our ‘sequence-ascertained favorable editing’ (SAFE) donor approach relies on the use of DNA donor mixtures containing the desired nucleotide substitutions or the wild-type alleles together with combinations of additional ‘diagnostic’ substitutions unlikely to have any effects. Sequencing of the target sites then results in that two different sequences are seen when both chromosomes are edited with ‘SAFE’ donors containing different sets of substitutions, while a single sequence indicates unintended effects such as deletions or gene conversion. We analyzed more than 850 human embryonic stem cell clones edited with ‘SAFE’ donors and detect all copy number changes and almost all clones with gene conversion. | |||||
出版者 | ||||||
出版者 | Oxford University Press | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 0305-1048 | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 1362-4962 | |||||
PubMed番号 | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | PMID | |||||
関連識別子 | info:pmid/36620901 | |||||
DOI | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | DOI | |||||
関連識別子 | info:doi/10.1093/nar/gkac1254 | |||||
権利 | ||||||
権利情報 | © 2023 The Authors. | |||||
関連サイト | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://academic.oup.com/nar/article/51/5/e26/6976061?login=true | |||||
著者版フラグ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 |