Data from: Pooled genome-wide CRISPR activation screening for rapamycin resistance genes in Drosophila cells
Cite this dataset
Xia, Baolong et al. (2023). Data from: Pooled genome-wide CRISPR activation screening for rapamycin resistance genes in Drosophila cells [Dataset]. Dryad. https://doi.org/10.5061/dryad.2547d7ww8
Loss-of-function and gain-of-function genetic perturbations provide valuable insights into gene function. In Drosophila cells, while genome-wide loss-of-function screens have been extensively used to reveal mechanisms of a variety of biological processes, approaches for performing genome-wide gain-of-function screens are still lacking. Here, we describe a pooled CRISPR activation (CRISPRa) screening platform in Drosophila cells and apply this method to both focused and genome-wide screens to identify rapamycin resistance genes. The screens identified three genes as novel rapamycin resistance genes: a member of SLC16 family of monocarboxylate transporters (CG8468), a member of the lipocalin protein family (CG5399), and a zinc finger C2H2 transcription factor (CG9932). Mechanistically, we demonstrate that CG5399 overexpression activates the RTK-Akt-mTOR signaling pathway and that activation of insulin receptor (InR) by CG5399 requires cholesterol and clathrin-coated pits at the cell membrane. This study establishes a novel platform for functional genetic studies in Drosophila cells.
The library was sequenced using NextSeq500 1 × 75 SE platform (Illumina) in HMS Biopolymers Facility. The sequencing data were de-multiplexed using TagDust. Screen hits were identified using MAGeCK-RRA.
Screen results are analyzed using MAGeCK-RRA. Excel can be used to open the data files.
National Institute of General Medical Sciences, Award: GM132087
National Cancer Institute, Award: CA120964
Howard Hughes Medical Institute