Nature research paper: CRISPR screens in Drosophila cells identify Vsg as a Tc toxin receptor
, Prevalence of all sgRNAs targeting Vsg, Mop, FIG4, or VAC14 across three rounds of pTc or mTc screens were quantified and plotted according to their corresponding gRNA abundances., Schematic of the experimental steps. Briefly, S2 cells were bathed in dsRNAs targeting the top hits from pTc CRISPR screen # 1 round 1 and allowed to recover for 3 days. Next, cells were treated with 100 pM pTc for 2 days, re-seeded and grown for an additional 7 days.
Gate representing DNA content of 80–90% of untreated cells was set. There was no pre-gating on cell sizes as pTc treatment causes changes in cell sizes and shapes., The indicated top hits from pTc CRISPR screens were knocked down in S2 cells using dsRNA and then exposed to pTc as described in . Percentage of cells with normal Hoechst levels were plotted., Comparison of untreated cells versus those treated with 100 pM pTc revealed increase in cell size .
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