Cargando…
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...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cell Press
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3714591/ https://www.ncbi.nlm.nih.gov/pubmed/23827738 http://dx.doi.org/10.1016/j.celrep.2013.06.020 |
_version_ | 1782277382471680000 |
---|---|
author | Bassett, Andrew R. Tibbit, Charlotte Ponting, Chris P. Liu, Ji-Long |
author_facet | Bassett, Andrew R. Tibbit, Charlotte Ponting, Chris P. Liu, Ji-Long |
author_sort | Bassett, Andrew R. |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-3714591 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Cell Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-37145912013-07-18 Highly Efficient Targeted Mutagenesis of Drosophila with the CRISPR/Cas9 System Bassett, Andrew R. Tibbit, Charlotte Ponting, Chris P. Liu, Ji-Long Cell Rep Resource 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. Cell Press 2013-07-11 /pmc/articles/PMC3714591/ /pubmed/23827738 http://dx.doi.org/10.1016/j.celrep.2013.06.020 Text en © 2013 The Authors https://creativecommons.org/licenses/by-nc-nd/3.0/ Open Access under CC BY-NC-ND 3.0 (https://creativecommons.org/licenses/by-nc-nd/3.0/) license |
spellingShingle | Resource Bassett, Andrew R. Tibbit, Charlotte Ponting, Chris P. Liu, Ji-Long 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 |
topic | Resource |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3714591/ https://www.ncbi.nlm.nih.gov/pubmed/23827738 http://dx.doi.org/10.1016/j.celrep.2013.06.020 |
work_keys_str_mv | AT bassettandrewr highlyefficienttargetedmutagenesisofdrosophilawiththecrisprcas9system AT tibbitcharlotte highlyefficienttargetedmutagenesisofdrosophilawiththecrisprcas9system AT pontingchrisp highlyefficienttargetedmutagenesisofdrosophilawiththecrisprcas9system AT liujilong highlyefficienttargetedmutagenesisofdrosophilawiththecrisprcas9system |