Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase

The CRISPR/Cas9 nickase mutant is less prone to off-target double-strand (ds)DNA breaks than wild-type Cas9 because to produce dsDNA cleavage it requires two guide RNAs to target the nickase to nearby opposing strands. Like wild-type Cas9 lesions, these staggered lesions are repaired by either non-h...

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Detalles Bibliográficos
Autores principales: Ge, Xi A., Hunter, Craig P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Genetics Society of America 2018
Materias:
Investigations
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6325892/
https://www.ncbi.nlm.nih.gov/pubmed/30504134
http://dx.doi.org/10.1534/g3.118.200758
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author Ge, Xi A.
Hunter, Craig P.
author_facet Ge, Xi A.
Hunter, Craig P.
author_sort Ge, Xi A.
collection PubMed
description The CRISPR/Cas9 nickase mutant is less prone to off-target double-strand (ds)DNA breaks than wild-type Cas9 because to produce dsDNA cleavage it requires two guide RNAs to target the nickase to nearby opposing strands. Like wild-type Cas9 lesions, these staggered lesions are repaired by either non-homologous end joining or, if a repair template is provided, by homologous recombination (HR). Here, we report very efficient (up to 100%) recovery of heterozygous insertions in Mus musculus produced by long (>300 nt), single-stranded DNA donor template-guided repair of paired-nickase lesions.
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spelling pubmed-63258922019-01-10 Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase Ge, Xi A. Hunter, Craig P. G3 (Bethesda) Investigations The CRISPR/Cas9 nickase mutant is less prone to off-target double-strand (ds)DNA breaks than wild-type Cas9 because to produce dsDNA cleavage it requires two guide RNAs to target the nickase to nearby opposing strands. Like wild-type Cas9 lesions, these staggered lesions are repaired by either non-homologous end joining or, if a repair template is provided, by homologous recombination (HR). Here, we report very efficient (up to 100%) recovery of heterozygous insertions in Mus musculus produced by long (>300 nt), single-stranded DNA donor template-guided repair of paired-nickase lesions. Genetics Society of America 2018-11-21 /pmc/articles/PMC6325892/ /pubmed/30504134 http://dx.doi.org/10.1534/g3.118.200758 Text en Copyright © 2019 by the Genetics Society of America http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigations
Ge, Xi A.
Hunter, Craig P.
Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase
title Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase
title_full Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase
title_fullStr Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase
title_full_unstemmed Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase
title_short Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase
title_sort efficient homologous recombination in mice using long single stranded dna and crispr cas9 nickase
topic Investigations
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6325892/
https://www.ncbi.nlm.nih.gov/pubmed/30504134
http://dx.doi.org/10.1534/g3.118.200758
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