| The Gene Disruption Project (GDP) is using non-targeted transposon mutagenesis and homologous recombination based targeted insertion strategies to create a public collection of insertion mutants. The collection can be searched online and new mutants may be requested while they are being balanced. Balanced stocks are available from the Bloomington Drosophila Stock Center. We have currently contributed over 25,000 insertions to the public collection. |
| Current Phase |
| In the current phase of our project, we are creating CRIMIC lines to generate GAL4 gene trap alleles. We developed novel methods to be able to use commercial gene synthesis for cloning homology donor constructs with short homology arms (see the CRIMIC section). The GDP project is currently supported by the Office of Research Infrastructure Programs (ORIP) (R24OD031447). The CRIMIC production pipeline is led by Oguz Kanca with help of Megan Cooper and Catherine Grace Burns (cloning), Wen-wen Lin, Ying Fang, Junyan Fang (injections), and Xinhua Huang and Minghua Qin Idso (stock creations) and Ming Ge and Ruifang Zhang (quality control) in the Bellen lab. Bob Levis (Carngie, Allan Spradling lab) performs the manual curation of the data and submission of the data to FlyBase and BDSC. The teams have generated more than 3500 GAL4 strains so far and the stocks have been distributed to nearly 500 labs in 36 countries in 2024. |
| Previous Phases |
| Phase I |
| This project was a collaboration among the laboratories of Hugo Bellen (Baylor College of Medicine), Roger Hoskins (Lawrence Berkeley National Laboratory) and Allan Spradling (Carnegie Institution of Washington), supported by the National Institutes of Health (R01GM067858) and the Howard Hughes Medical Institute. The flies were generated, selected, balanced and maintained by Yuchun He, Ying Fang, Zhihua Wang and Jianping Li in the Bellen lab. The inverse PCRs and sequencing were performed by Martha Evans-Holm in the Hoskins lab. Mapping information was obtained by Ben Booth in the Hoskins lab followed by manual curation by Bob Levis and Allan Spradling in the Spradling lab. A variety of transposons were used as we continued to upgrade to strive for better coverage, and we also sequenced and selected insertions from donated collections for deposition in the BDSC. |
| Phase II |
| During this part of the project, the MiMIC insertion was developed in the Bellen lab by Koen Venken. About ~17,500 MiMIC insertions were generated at Baylor College of Medicine by the Bellen Lab team and were end-sequenced by the Hoskins lab at Lawrence Berkeley National Laboratory. The sequence information was curated by Bob Levis at Carnegie Institution for Science. A subset of these insertions were selected for RMCE conversion using the GFSTF and TG4 tags using both the injection and crossing methods. |
| Phase III |
| In phase III of our project we started using CRISPR/Cas9 mediated homologous recombination to create CRIMIC lines. This project started as a collaboration among the laboratories of Hugo Bellen (Baylor College of Medicine), Norbert Perrimon (Harvard Medical School) and Allan Spradling (Carnegie Institution for Science) and was supported by the National Institutes of Health (R01GM067858) and the Howard Hughes Medical Institute. The Perrimon lab designed and cloned the CRIMIC constructs (Jonathan Zirin, Yanhui Hu, Stephanie Mohr, Rong Tao and Colby Devereaux); new method development, injection, balancing and quality control checks were performed in the Bellen Lab (Oguz Kanca, Wen-wen Lin, Yuchun He, Ying Fang, Qiaohong Gao, Liwen Ma, Junyan Fang, Zhihua Wang, Ming Ge and Jiangxing Lv) and stock data transfer to FlyBase and BDSC was handled by Bob Levis (Carnegie). |
| Please reference our publications, which are also found on the Downloads page, as well as the Bloomington Drosophila Stock Center, when utilizing these stocks. References
|