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: | Article |
| Language: | English |
| Published: |
Elsevier
2013-07-01
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| Series: | Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124713003124 |
| _version_ | 1811319483518681088 |
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| author | Andrew R. Bassett Charlotte Tibbit Chris P. Ponting Ji-Long Liu |
| author_facet | Andrew R. Bassett Charlotte Tibbit Chris P. Ponting Ji-Long Liu |
| author_sort | Andrew R. Bassett |
| collection | DOAJ |
| 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 | 2024-04-13T12:43:57Z |
| format | Article |
| id | doaj.art-b1c976c5d486437cb21d984cf3d133bf |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-04-13T12:43:57Z |
| publishDate | 2013-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-b1c976c5d486437cb21d984cf3d133bf2022-12-22T02:46:26ZengElsevierCell Reports2211-12472013-07-014122022810.1016/j.celrep.2013.06.020Highly Efficient Targeted Mutagenesis of Drosophila with the CRISPR/Cas9 SystemAndrew R. Bassett0Charlotte Tibbit1Chris P. Ponting2Ji-Long Liu3Medical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3PT, UKMedical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3PT, UKMedical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3PT, UKMedical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3PT, UKHere, 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.http://www.sciencedirect.com/science/article/pii/S2211124713003124 |
| spellingShingle | Andrew R. Bassett Charlotte Tibbit Chris P. Ponting Ji-Long Liu Highly Efficient Targeted Mutagenesis of Drosophila with the CRISPR/Cas9 System Cell Reports |
| 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 |
| url | http://www.sciencedirect.com/science/article/pii/S2211124713003124 |
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