Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring
The emergence of the CRISPR-Cas9 system has triggered a technical revolution in mammalian genome editing. Compared to traditional gene-targeting strategies, CRISPR-Cas9 technology offers a more efficient and cost-effective approach for generating genetically modified animal models. However, off-targ...
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Oxford University Press
2019-11-01
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Online Access: | http://g3journal.org/lookup/doi/10.1534/g3.119.400503 |
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doaj-99edfd2f5c7045f894d55f538d1aa3842021-07-02T07:58:46ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362019-11-019113645365110.1534/g3.119.40050315Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their OffspringYan DongHaimei LiLiang ZhaoPeter KoopmanFeng ZhangJohnny X. HuangThe emergence of the CRISPR-Cas9 system has triggered a technical revolution in mammalian genome editing. Compared to traditional gene-targeting strategies, CRISPR-Cas9 technology offers a more efficient and cost-effective approach for generating genetically modified animal models. However, off-target cleavage in CRISPR-mediated genome editing is a major concern in the analysis of phenotypes as well as the selection of therapeutic targets. Here, we analyzed whole-genome sequencing (WGS) data from two knock-out (KO) mouse strains generated by using the CRISPR-Cas9 system targeting the Mmd and Paqr8 loci. A total of nine individuals were sequenced including two parents, four F1 offspring and three uninjected control mice. Using GATK and bcftools software, we identified two off-target events in the founder mice. The two CRISPR-Cas9-induced off-target events were predictable using Cas-OFFinder and were not passed on to the offspring that we investigated. In addition, our results indicated that the number of CRISPR-Cas9-induced mutations was not statistically distinguishable from the background de novo mutations (DNMs).http://g3journal.org/lookup/doi/10.1534/g3.119.400503crispr-cas9off-targetwhole-genome sequencinggenome editing |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yan Dong Haimei Li Liang Zhao Peter Koopman Feng Zhang Johnny X. Huang |
spellingShingle |
Yan Dong Haimei Li Liang Zhao Peter Koopman Feng Zhang Johnny X. Huang Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring G3: Genes, Genomes, Genetics crispr-cas9 off-target whole-genome sequencing genome editing |
author_facet |
Yan Dong Haimei Li Liang Zhao Peter Koopman Feng Zhang Johnny X. Huang |
author_sort |
Yan Dong |
title |
Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring |
title_short |
Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring |
title_full |
Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring |
title_fullStr |
Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring |
title_full_unstemmed |
Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring |
title_sort |
genome-wide off-target analysis in crispr-cas9 modified mice and their offspring |
publisher |
Oxford University Press |
series |
G3: Genes, Genomes, Genetics |
issn |
2160-1836 |
publishDate |
2019-11-01 |
description |
The emergence of the CRISPR-Cas9 system has triggered a technical revolution in mammalian genome editing. Compared to traditional gene-targeting strategies, CRISPR-Cas9 technology offers a more efficient and cost-effective approach for generating genetically modified animal models. However, off-target cleavage in CRISPR-mediated genome editing is a major concern in the analysis of phenotypes as well as the selection of therapeutic targets. Here, we analyzed whole-genome sequencing (WGS) data from two knock-out (KO) mouse strains generated by using the CRISPR-Cas9 system targeting the Mmd and Paqr8 loci. A total of nine individuals were sequenced including two parents, four F1 offspring and three uninjected control mice. Using GATK and bcftools software, we identified two off-target events in the founder mice. The two CRISPR-Cas9-induced off-target events were predictable using Cas-OFFinder and were not passed on to the offspring that we investigated. In addition, our results indicated that the number of CRISPR-Cas9-induced mutations was not statistically distinguishable from the background de novo mutations (DNMs). |
topic |
crispr-cas9 off-target whole-genome sequencing genome editing |
url |
http://g3journal.org/lookup/doi/10.1534/g3.119.400503 |
work_keys_str_mv |
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1721335370896900096 |