Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system
Here, we present a simple and highly efficient method for generating and detecting mutations of any gene in Drosophila melanogaster through the use of the CRISPR/Cas9 system (clustered regularly interspaced palindromic repeats/CRISPR-associated). We show that injection of RNA into the Drosophila emb...
| Main Authors: | , , , |
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| Format: | Journal article |
| Language: | English |
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Cell Press
2013
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| _version_ | 1797111719999307776 |
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| author | Bassett, AR Tibbit, C Ponting, CP Liu, J-L |
| author_facet | Bassett, AR Tibbit, C Ponting, CP Liu, J-L |
| author_sort | Bassett, AR |
| collection | OXFORD |
| description | Here, we present a simple and highly efficient method for generating and detecting mutations of any gene in Drosophila melanogaster through the use of the CRISPR/Cas9 system (clustered regularly interspaced palindromic repeats/CRISPR-associated). We show that injection of RNA into the Drosophila embryo can induce highly efficient mutagenesis of desired target genes in up to 88% of injected flies. These mutations can be transmitted through the germline to make stable lines. Our system provides at least a 10-fold improvement in efficiency over previously published reports, enabling wider application of this technique. We also describe a simple and highly sensitive method of detecting mutations in the target gene by high-resolution melt analysis and discuss how the new technology enables the study of gene function. |
| first_indexed | 2024-03-07T08:14:15Z |
| format | Journal article |
| id | oxford-uuid:8d139179-ac13-492e-8632-2b93002f7cb4 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T08:14:15Z |
| publishDate | 2013 |
| publisher | Cell Press |
| record_format | dspace |
| spelling | oxford-uuid:8d139179-ac13-492e-8632-2b93002f7cb42023-12-13T12:00:44ZHighly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 systemJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:8d139179-ac13-492e-8632-2b93002f7cb4EnglishSymplectic Elements at OxfordCell Press2013Bassett, ARTibbit, CPonting, CPLiu, J-LHere, we present a simple and highly efficient method for generating and detecting mutations of any gene in Drosophila melanogaster through the use of the CRISPR/Cas9 system (clustered regularly interspaced palindromic repeats/CRISPR-associated). We show that injection of RNA into the Drosophila embryo can induce highly efficient mutagenesis of desired target genes in up to 88% of injected flies. These mutations can be transmitted through the germline to make stable lines. Our system provides at least a 10-fold improvement in efficiency over previously published reports, enabling wider application of this technique. We also describe a simple and highly sensitive method of detecting mutations in the target gene by high-resolution melt analysis and discuss how the new technology enables the study of gene function. |
| spellingShingle | Bassett, AR Tibbit, C Ponting, CP Liu, J-L Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system |
| title | Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system |
| title_full | Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system |
| title_fullStr | Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system |
| title_full_unstemmed | Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system |
| title_short | Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system |
| title_sort | highly efficient targeted mutagenesis of drosophila with the crispr cas9 system |
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