eLifeGeneticsPeer-reviewed journal article
Efficient single-copy HDR by 5’ modified long dsDNA donors
José Arturo Gutierrez-Triana, Tinatini TavhelidseiD, Thomas ThumbergeriD, Isabelle Thomas, Beate WittbrodtiD, Tanja Kellner, Kerim AnlaşiD, Erika TsingosiD, Joachim WittbrodtiD
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.
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- top 7%
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