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Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila
Open accessPublished version of a preprint
Abstract
Significance: Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas)-mediated genome engineering promises to revolutionize genetic studies in a variety of systems. Here we describe an optimized set of tools for CRISPR/Cas experiments in the model organism Drosophila melanogaster . These tools can be used for remarkably efficient germline transmission of ( i ) loss-of-function insertion and deletion mutations in essential genes or ( ii ) precise changes in the genome sequence introduced by homology-directed repair. These tools also permit efficient biallelic targeting of genes in somatic cells, thereby demonstrating a novel application of CRISPR/Cas in rapidly revealing mutant phenotypes within the organism. Our work also paves the way for high-throughput genetic screens in Drosophila with CRISPR/Cas.
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Metadata
- Authors
- Port, F., Chen, H.-M., Lee, T., & Bullock, S. L.
- Affiliations
- MRC Laboratory of Molecular Biology (GB)
- Howard Hughes Medical Institute (US)
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2014-07-07
- Volume
- 111
- Issue
- 29
- Pages
- E2967-76
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC4115528
- OpenAlex
- W1998356880
- Cited by
- 1,046
- References
- 39
- Citations by year
- 2026: 57 · 2025: 73 · 2024: 93 · 2023: 98 · 2022: 108 · 2021: 113 · 2020: 125 · 2019: 104 · 2018: 88 · 2017: 67 · 2016: 58 · 2015: 48 · 2014: 13
- FWCI
- 25.44
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- cc-by-nc-nd
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- The Journal of the American Medical Association (JAMA) Network (American Medical Association) (submittedVersion, other-oa)
- Funders
- Howard Hughes Medical Institute
- Medical Research Council (U105178790)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Keywords
- CRISPR/Cas9, Drosophila melanogaster, germline editing, somatic genome editing, homology-directed repair, loss-of-function phenotypes, high-throughput genetic screening
- MeSH terms
- Alleles, Animals, Base Sequence, DNA Repair, Drosophila melanogaster, Genetic Engineering, Genetic Vectors, Germ Cells, Molecular Sequence Data, Mutation, Phenotype, RNA, Genes, Insect, Gene Targeting, Transgenes, Genes, Essential, Animals, Genetically Modified, Genome, Insect, CRISPR-Cas Systems
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