Efficient single-copy HDR by 5' modified long dsDNA donors
- Heidelberg University, Germany
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.
Data availability
All data generated or analyzed during this study are included in the manuscript and supporting files. Source data files have been provided for Figure 2-figure supplement 1.
Article and author information
Author details
Tinatini Tavhelidse
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
Version 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
© 2018, Gutierrez-Triana et al.
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.
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Efficient single-copy HDR by 5' modified long dsDNA donors
eLife 7:e39468.
https://doi.org/10.7554/eLife.39468
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