The Gene Disruption Project (GDP) has provided the research community with new genetic tools to support the functional analysis of the Drosophila genome and is responsible for generating 85% of the more than 15,000 transposable element (TE) insertion mutant stocks that are present in the Bloomington Drosophila Stock Center (BDSC). The newest stocks were generated using MiMIC, a TE that allows site-specific recombination-based tools for in vivo manipulation of the genome. We now propose to expand the GDP collection to increase its coverage and provide new methods for analyzing gene function. With the advent of the CRISPR gene editing technology, it is now technically feasible to make targeted modifications in user-defined locations rather than at random as is done with TEs. We propose to use CRISPR to introduce swappable insertion cassettes (SICs) into 5,000 genes missed by MiMIC. Three thousand genes that are evolutionarily conserved and are likely to be implicated in human disease will be prioritized. The remaining 2,000 will be selected based on the needs and interests of the community. In addition, we propose to generate protein trap alleles for 2,500 genes using existing MiMIC lines as well as new insertion lines developed in this proposal. These protein trap alleles will encode an in-frame GFP fusion and allow the determination of precise protein distribution using light and electron microscopy. They also allow efficient knockdown of mRNA transcript or protein product in specific cells or tissues using RNAi and deGradFP methodologies against the GFP tag. Furthermore, protein tagging allows purification strategies using nanobodies against GFP such as immunoprecipitation (IP) of proteins, chromatin IP for DNA-associated proteins, and IP-mass spectroscopy. New insertion and protein trap strains will be validated, and the supporting information submitted to public repositories such as GenBank and Flybase. Stocks generated by the GDP will be deposited at the BDSC for public distribution.

Public Health Relevance

Research with model organisms such as the fruit fly, Drosophila melanogaster, has given us a much better understanding of the biology of higher organisms, including humans, due to the striking evolutionary conservation of gene function. Many genes discovered in Drosophila have been implicated in human diseases such as cancer, neurological disorders, metabolic disorders, and developmental diseases. By generating additional mutant strains and tools described in this proposal, we are providing valuable resources that will greatly advance the pace of basic and translational research in many laboratories around the world.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
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Genomics, Computational Biology and Technology Study Section (GCAT)
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Willis, Kristine Amalee
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Baylor College of Medicine
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Li-Kroeger, David; Kanca, Oguz; Lee, Pei-Tseng et al. (2018) An expanded toolkit for gene tagging based on MiMIC and scarless CRISPR tagging in Drosophila. Elife 7:
?entürk, Mümine; Bellen, Hugo J (2018) Genetic strategies to tackle neurological diseases in fruit flies. Curr Opin Neurobiol 50:24-32
Marcogliese, Paul C; Shashi, Vandana; Spillmann, Rebecca C et al. (2018) IRF2BPL Is Associated with Neurological Phenotypes. Am J Hum Genet 103:245-260
Tan, Kai Li; Haelterman, Nele A; Kwartler, Callie S et al. (2018) Ari-1 Regulates Myonuclear Organization Together with Parkin and Is Associated with Aortic Aneurysms. Dev Cell 45:226-244.e8
Cosmanescu, Filip; Katsamba, Phinikoula S; Sergeeva, Alina P et al. (2018) Neuron-Subtype-Specific Expression, Interaction Affinities, and Specificity Determinants of DIP/Dpr Cell Recognition Proteins. Neuron 100:1385-1400.e6
Liu, Ning; Schoch, Kelly; Luo, Xi et al. (2018) Functional variants in TBX2 are associated with a syndromic cardiovascular and skeletal developmental disorder. Hum Mol Genet 27:2454-2465
Lee, Pei-Tseng; Lin, Guang; Lin, Wen-Wen et al. (2018) A kinase-dependent feedforward loop affects CREBB stability and long term memory formation. Elife 7:
Chao, Hsiao-Tuan; Davids, Mariska; Burke, Elizabeth et al. (2017) A Syndromic Neurodevelopmental Disorder Caused by De Novo Variants in EBF3. Am J Hum Genet 100:128-137
Wangler, Michael F; Yamamoto, Shinya; Chao, Hsiao-Tuan et al. (2017) Model Organisms Facilitate Rare Disease Diagnosis and Therapeutic Research. Genetics 207:9-27
Luo, Xi; Rosenfeld, Jill A; Yamamoto, Shinya et al. (2017) Clinically severe CACNA1A alleles affect synaptic function and neurodegeneration differentially. PLoS Genet 13:e1006905

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