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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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
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author Gutierrez-Triana, Jose Arturo
Tavhelidse, Tinatini
Thumberger, Thomas
Thomas, Isabelle
Wittbrodt, Beate
Kellner, Tanja
Anlas, Kerim
Tsingos, Erika
Wittbrodt, Joachim
author_facet Gutierrez-Triana, Jose Arturo
Tavhelidse, Tinatini
Thumberger, Thomas
Thomas, Isabelle
Wittbrodt, Beate
Kellner, Tanja
Anlas, Kerim
Tsingos, Erika
Wittbrodt, Joachim
author_sort Gutierrez-Triana, Jose Arturo
collection PubMed
description 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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spelling pubmed-61251272018-09-07 Efficient single-copy HDR by 5’ modified long dsDNA donors Gutierrez-Triana, Jose Arturo Tavhelidse, Tinatini Thumberger, Thomas Thomas, Isabelle Wittbrodt, Beate Kellner, Tanja Anlas, Kerim Tsingos, Erika Wittbrodt, Joachim eLife Developmental Biology 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. eLife Sciences Publications, Ltd 2018-08-29 /pmc/articles/PMC6125127/ /pubmed/30156184 http://dx.doi.org/10.7554/eLife.39468 Text en © 2018, Gutierrez-Triana et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Developmental Biology
Gutierrez-Triana, Jose Arturo
Tavhelidse, Tinatini
Thumberger, Thomas
Thomas, Isabelle
Wittbrodt, Beate
Kellner, Tanja
Anlas, Kerim
Tsingos, Erika
Wittbrodt, Joachim
Efficient single-copy HDR by 5’ modified long dsDNA donors
title Efficient single-copy HDR by 5’ modified long dsDNA donors
title_full Efficient single-copy HDR by 5’ modified long dsDNA donors
title_fullStr Efficient single-copy HDR by 5’ modified long dsDNA donors
title_full_unstemmed Efficient single-copy HDR by 5’ modified long dsDNA donors
title_short Efficient single-copy HDR by 5’ modified long dsDNA donors
title_sort efficient single-copy hdr by 5’ modified long dsdna donors
topic Developmental Biology
url 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
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