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

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Cite this article as: eLife 2018;7:e39468 doi: 10.7554/eLife.39468

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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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Jose Arturo Gutierrez-Triana

Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany

Competing interests
The authors declare that no competing interests exist.
Tinatini Tavhelidse

Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany

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The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-6103-9019
Thomas Thumberger

Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany

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The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-8485-457X
Isabelle Thomas

Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany

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The authors declare that no competing interests exist.
Beate Wittbrodt

Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany

Competing interests
The authors declare that no competing interests exist.
Tanja Kellner

Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany

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The authors declare that no competing interests exist.
Kerim Anlas

Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany

Competing interests
The authors declare that no competing interests exist.
Erika Tsingos

Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany

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The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-7267-160X
Joachim Wittbrodt

Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany

For correspondence
jochen.wittbrodt@cos.uni-heidelberg.de

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-8550-7377

Funding

Deutsche Forschungsgemeinschaft (CRC 873 project A3)

Joachim Wittbrodt

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

Alejandro Sánchez Alvarado, Stowers Institute for Medical Research, United States

Publication history

Received: June 22, 2018
Accepted: August 14, 2018
Accepted Manuscript published: August 29, 2018 (version 1)
Version of Record published: September 5, 2018 (version 2)

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.