Efficient Single-copy HDR by 5' Modified Long DsDNA Donors

Overview

Journal Elife

Specialty Biology

Date 2018 Aug 30

PMID 30156184

Citations 55

Authors

Jose Arturo Gutierrez-Triana

Tinatini Tavhelidse

Thomas Thumberger

Isabelle Thomas

Beate Wittbrodt

Tanja Kellner

Kerim Anlas

Erika Tsingos

Joachim Wittbrodt

Affiliations

Soon will be listed here.

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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