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...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Oxford University Press
2019-11-01
|
| Series: | G3: Genes, Genomes, Genetics |
| Subjects: | |
| Online Access: | http://g3journal.org/lookup/doi/10.1534/g3.119.400503 |
| _version_ | 1818623081748365312 |
|---|---|
| author | Yan Dong Haimei Li Liang Zhao Peter Koopman Feng Zhang Johnny X. Huang |
| author_facet | Yan Dong Haimei Li Liang Zhao Peter Koopman Feng Zhang Johnny X. Huang |
| author_sort | Yan Dong |
| collection | DOAJ |
| 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). |
| first_indexed | 2024-12-16T18:35:24Z |
| format | Article |
| id | doaj.art-99edfd2f5c7045f894d55f538d1aa384 |
| institution | Directory Open Access Journal |
| issn | 2160-1836 |
| language | English |
| last_indexed | 2024-12-16T18:35:24Z |
| publishDate | 2019-11-01 |
| publisher | Oxford University Press |
| record_format | Article |
| series | G3: Genes, Genomes, Genetics |
| spelling | doaj.art-99edfd2f5c7045f894d55f538d1aa3842022-12-21T22:21:10ZengOxford 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 |
| 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 |
| title | 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_short | 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 |
| 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 | AT yandong genomewideofftargetanalysisincrisprcas9modifiedmiceandtheiroffspring AT haimeili genomewideofftargetanalysisincrisprcas9modifiedmiceandtheiroffspring AT liangzhao genomewideofftargetanalysisincrisprcas9modifiedmiceandtheiroffspring AT peterkoopman genomewideofftargetanalysisincrisprcas9modifiedmiceandtheiroffspring AT fengzhang genomewideofftargetanalysisincrisprcas9modifiedmiceandtheiroffspring AT johnnyxhuang genomewideofftargetanalysisincrisprcas9modifiedmiceandtheiroffspring |