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

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 w...

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Main Authors: Gutiérrez Triana, José Arturo (Author) , Tavhelidse-Suck, Tinatini (Author) , Thumberger, Thomas (Author) , Thomas, Isabelle (Author) , Wittbrodt, Beate (Author) , Kellner, Tanja (Author) , Anlas, Kerim (Author) , Tsingos, Erika (Author) , Wittbrodt, Joachim (Author)
Format: Article (Journal)
Language:English
Published: Aug 29, 2018
In: eLife
Year: 2018, Volume: 7
ISSN:2050-084X
DOI:10.7554/eLife.39468
Online Access:Resolving-System, kostenfrei, Volltext: http://dx.doi.org/10.7554/eLife.39468
Verlag, kostenfrei, Volltext: https://elifesciences.org/articles/39468
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Author Notes:Jose Arturo Gutierrez-Triana, Tinatini Tavhelidse, Thomas Thumberger, Isabelle Thomas, Beate Wittbrodt, Tanja Kellner, Kerim Anlas, Erika Tsingos and Joachim Wittbrodt
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Summary: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.
Item Description:Gesehen am 04.09.2018
Physical Description:Online Resource
ISSN:2050-084X
DOI:10.7554/eLife.39468

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