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Efficient single-copy HDR by 5' modified long dsDNA donors.

ABSTRACT: CRISPR/Cas9 efficiently induces targeted mutations via non-homologous-end-joining but for genome editing, precise, homology-directed repair (HDR) of endogenous DNA stretches is a prerequisite. To favor HDR, many approaches interfere with the repair machinery or manipulate Cas9 itself. Using Medaka we show that the modification of 5' ends of long dsDNA donors strongly enhances HDR, favors efficient single-copy integration by retaining a monomeric donor conformation thus facilitating successful gene replacement or tagging.

SUBMITTER: Gutierrez-Triana JA

PROVIDER: S-EPMC6125127 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature

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Efficient single-copy HDR by 5' modified long dsDNA donors.

Gutierrez-Triana Jose Arturo JA Tavhelidse Tinatini T Thumberger Thomas T Thomas Isabelle I Wittbrodt Beate B Kellner Tanja T Anlas Kerim K Tsingos Erika E Wittbrodt Joachim J

eLife 20180829

CRISPR/Cas9 efficiently induces targeted mutations via non-homologous-end-joining but for genome editing, precise, homology-directed repair (HDR) of endogenous DNA stretches is a prerequisite. To favor HDR, many approaches interfere with the repair machinery or manipulate Cas9 itself. Using Medaka we show that the modification of 5' ends of long dsDNA donors strongly enhances HDR, favors efficient single-copy integration by retaining a monomeric donor conformation thus facilitating successful ge ...[more]

PMID: 30156184

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ssDNA is not superior to dsDNA as long HDR donors for CRISPR-mediated endogenous gene tagging in human diploid RPE1 and HCT116 cells.

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Author Correction: SMURF-seq: efficient copy number profiling on long-read sequencers.

Project description:An amendment to this paper has been published and can be accessed via the original article.

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