Pooled Genome-Wide CRISPR Screening for Basal and Context-Specific Fitness Gene Essentiality in Drosophila Cells
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CitationViswanatha, Raghuvir, Zhongchi Li, Yanhui Hu, Norbert Perrimon. "Pooled Genome-Wide CRISPR Screening for Basal and Context-Specific Fitness Gene Essentiality in Drosophila Cells." eLife 7 (2018): e36333. DOI: 10.7554/elife.36333
AbstractGenome-wide screens in Drosophila cells have offered numerous insights into gene function, yet a major limitation has been the inability to stably deliver large multiplexed DNA libraries to cultured cells allowing barcoded pooled screens. Here, we developed a site-specific integration strategy for library delivery and performed a genome-wide CRISPR knockout screen in Drosophila S2R+ cells. Under basal growth conditions, 1235 genes were essential for cell fitness at a false-discovery rate of 5%, representing the highest-resolution fitness gene set yet assembled for Drosophila, including 407 genes which likely duplicated along the vertebrate lineage and whose orthologs were underrepresented in human CRISPR screens. We additionally performed context-specific fitness screens for resistance to or synergy with trametinib, a Ras/ERK/ETS inhibitor, or rapamycin, an mTOR inhibitor, and identified key regulators of each pathway. The results present a novel, scalable, and versatile platform for functional genomic screens in invertebrate cells.
Citable link to this pagehttps://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37369746
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