Genome-wide CRISPR Knockout Screen in HAP1 MTHFD1-K56R cells to identify genes which confer to resistance to adenosine toxicity

This dataset contains sequencing data from a pooled CRISPR knockout screen performed in HAP1 MTHFD1K56R (i.e. harbouring a K56R mutation in MTHFD1) cells. Cells were transduced with a lentiviral genome-wide sgRNA knockout library (Brunello, Addgene #73179) followed by puromycin selection. The screen was then performed in IMDM medium with dialysed FBS supplemented with 50µM adenosine (ADE). Under these conditions, HAP1 MTHFD1K56R usually die. Dialysed medium (DIA) was used as control conditions. Cells were expanded for 14 days and surviving cells were collected for subsequent sequencing analyses. The screen aimed to identify genes whose loss would confer to adenosine resistance in HAP1 MTHFD1-K56R cells. Sequencing was performed to identify sgRNA abundances between adenosine treatment and control. Data was analyzed using PinAPL-Py. Overall design: MTHFD1-K56R cells were transduced with Brunello sgRNA KO library (MOI approx. 1, >500x sgRNA coverage), selected for puromycin and cultured for 14days in total in dialysed media -+ 50µM adenosine. Experiments were performed in duplicates (2x DIA = CTRL, 2x ADE)