Cited by 1,046CRISPR and Genetic EngineeringRNA regulation and diseaseAnimal Genetics and Reproduction

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Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila

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Port, Fillip, Chen, Hui-Min, Lee, Tzumin, Bullock, Simon L.·2014·Proceedings of the National Academy of Sciences

DOI 10.1073/pnas.1405500111PMC PMC4115528

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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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Self-citation

Authors: Port, F., Chen, H.-M., Lee, T., & Bullock, S. L.
Affiliations: MRC Laboratory of Molecular Biology (GB)
Howard Hughes Medical Institute (US)
ORCID: Port, Fillip
Chen, Hui-Min
Lee, Tzumin
Bullock, Simon L.
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
DOI: 10.1073/pnas.1405500111
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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Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila

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