ArticleProceedings of the National Academy of Sciences of the United States of America2008
A powerful method combining homologous recombination and site-specific recombination for targeted mutagenesis in Drosophila.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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Who cites it
36 citing papers in PubMed, 66 citations in OpenAlex.
- A powerful and highly efficient PAI-mediated transgenesis approach in Drosophila.Nucleic acids research · 2025Article
- REV1 coordinates a multi-faceted tolerance response to DNA alkylation damage and prevents chromosome shattering in Drosophila melanogaster.PLoS genetics · 2024Article
- Demonstration of in vivo engineered tandem duplications of varying sizes using CRISPR and recombinases in Drosophila melanogaster.G3 (Bethesda, Md.) · 2023Article
- Scarless Modification of the Drosophila Genome Near Any Mapped attP Sites.Current protocols · 2023Article
- Taming active transposons at Drosophila telomeres: The interconnection between HipHop's roles in capping and transcriptional silencing.PLoS genetics · 2021Article
- Redundant enhancers in the iab-5 domain cooperatively activate Abd-B in the A5 and A6 abdominal segments of Drosophila.Development (Cambridge, England) · 2021Article
- Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction.Journal of visualized experiments : JoVE · 2018Article
- Engineering a light-activated caspase-3 for precise ablation of neurons in vivo.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- MTV, an ssDNA Protecting Complex Essential for Transposon-Based Telomere Maintenance in Drosophila.PLoS genetics · 2016Article
- Enhanced specificity and efficiency of the CRISPR/Cas9 system with optimized sgRNA parameters in Drosophila.Cell reports · 2014Article
- Fast and efficient Drosophila melanogaster gene knock-ins using MiMIC transposons.G3 (Bethesda, Md.) · 2014Article
- Targeted genome engineering techniques in Drosophila.Methods (San Diego, Calif.) · 2014Review
- Coordination of transposon expression with DNA replication in the targeting of telomeric retrotransposons in Drosophila.The EMBO journal · 2014Article
- Generation of a useful roX1 allele by targeted gene conversion.G3 (Bethesda, Md.) · 2014Article
- Strategies for gene disruption in Drosophila.Cell & bioscience · 2014Review
- Optimized gene editing technology for Drosophila melanogaster using germ line-specific Cas9.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
- Article
- A Functional Comparison of the 3xP3 Promoter by Recombinase-Mediated Cassette Exchange in Drosophila and a Tephritid Fly, Anastrepha suspensa.G3 (Bethesda, Md.) · 2013Article
- Long-range targeted manipulation of the Drosophila genome by site-specific integration and recombinational resolution.Genetics · 2013Article
- Targeted gene replacement in Drosophila goes the distance.Genetics · 2013Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
Gene targeting provides a powerful tool for dissecting gene function. However, repeated targeting of a single locus remains a practice mostly limited to unicellular organisms that afford simple targeting methodologies. We developed an efficient method to repeatedly target a single locus in Drosophila. In this method, which we term "site-specific integrase mediated repeated targeting" (SIRT), an attP attachment site for the phage phiC31 integrase is first targeted to the vicinity of the gene of interest by homologous recombination. All subsequent modifications of that gene are introduced by phiC31-mediated integration of plasmids carrying an attB attachment site and the desired mutation. This highly efficient integration results in a tandem duplication of the target locus, which is then reduced into a single copy carrying the mutation, likely by the efficient "single strand annealing" mechanism, induced with a DNA double-strand break (DSB). We used SIRT to generate a series of six mutations in the Drosophila nbs gene, ranging from single amino acid replacements and small in-frame deletions to complete deletion of the gene. Because all of the components of SIRT are functional in many different organisms, it is readily adaptable to other multicellular organisms.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.