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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Detalles Bibliográficos
Autores principales: Gutierrez-Triana, Jose Arturo, Tavhelidse, Tinatini, Thumberger, Thomas, Thomas, Isabelle, Wittbrodt, Beate, Kellner, Tanja, Anlas, Kerim, Tsingos, Erika, Wittbrodt, Joachim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2018
Materias:
Developmental Biology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6125127/
https://www.ncbi.nlm.nih.gov/pubmed/30156184
http://dx.doi.org/10.7554/eLife.39468
Descripción
Sumario: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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