# A Comprehensive Resource for Manipulating the Drosophila Genome > **NIH NIH R24** · BAYLOR COLLEGE OF MEDICINE · 2022 · $401,067 ## Abstract PROJECT SUMMARY We propose to generate GFP protein trap alleles for 295 fly orthologs of human genes associated with AD based Genome Wide Association Studies (GWAS), AD genome sequencing efforts, and other bioinformatics tools. The parent grant for this supplement R24 GDP OD031447, supports the Drosophila Gene Disruption Project (GDP) that generates GAL4 gene trap alleles for genes that are orthologous to human genes. These GAL4 gene trap alleles create severe loss of function alleles of the targeted gene to allow phenotypic characterizations. They also allow to determine the expression pattern of the gene and can also be used in combination with UAS-cDNA transgenes to rescue the phenotypes induced by the GAL4 insertion. This allows us to assess rescue with the reference cDNA of the human orthologue as well as the variants found in patients. The GFP protein trap alleles that we propose to generate here are highly complementary to the GAL4 gene trap alleles and enable very different experimental strategies. The GFP protein trap alleles allow us to determine subcellular localization of the targeted gene product. Additionally, the GFP tagged protein can be used to immunopurify the protein together with its binding partners, allowing the identification of interacting proteins through Mass spectroscopy. In addition, we developed a strategy to remove the GFP protein in a temporal and tissue specific fashion that is reversible in any tissue or cell type based on the DeGradFp method and temperature shifts. Hence, creating a GFP protein trap allele to accompany every GAL4 gene trap allele very significantly increases the breadth of information we can obtain for each targeted gene. With this supplement, we will target a subset of the genes that we are generating GAL4 gene trap alleles, specifically 295 genes that have been proposed to be associated with AD with emphasis on the genes that are currently understudied. We will use CRISPR mediated homologous recombination to integrate GFP protein trap artificial exon in introns of genes that contain a suitable intron. For the genes where no such suitable intron can be found, we will replace either the entire coding sequence or exons where we want to integrate GFP tag depending on the size of the gene. We will employ the scarless DsRed dominant marker as a transgenic marker that can be removed from the final tagged alleles. We will inject the above constructs and verify the insertion site by genomic PCRs. Every generated fly line will be deposited to Bloomington Drosophila Stock Center (BDSC) and all the associated information will be readily available in our public web site and Flybase. We will select 20 GFP protein trap alleles for high priority genes where AD links are better established based on previous studies and determine the subcellular localization of these proteins by immunostaining. We then specifically remove the protein in adult animals using conditional GFP tagged protein removal using DegradFP an... ## Key facts - **NIH application ID:** 10498566 - **Project number:** 3R24OD031447-02S1 - **Recipient organization:** BAYLOR COLLEGE OF MEDICINE - **Principal Investigator:** HUGO J BELLEN - **Activity code:** R24 (R01, R21, SBIR, etc.) - **Funding institute:** NIH - **Fiscal year:** 2022 - **Award amount:** $401,067 - **Award type:** 3 - **Project period:** 2021-07-01 → 2025-03-31 ## Primary source NIH RePORTER: https://reporter.nih.gov/project-details/10498566 ## Citation > US National Institutes of Health, RePORTER application 10498566, A Comprehensive Resource for Manipulating the Drosophila Genome (3R24OD031447-02S1). Retrieved via AI Analytics 2026-05-29 from https://api.ai-analytics.org/grant/nih/10498566. Licensed CC0. --- *[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*