Genome-wide CRISPR Screen for resistance to dihydroartemisinin (DHA)

To elucidate which cellular signaling pathways are activated by DHA to induce cell death in mouse BCR-ABL+ B-ALL cells, an unbiased genetic screen was conducted in which Cas9-expressing mouse BCR-ABL+ B-ALL cells were stably transduced with the Brie knockout library (78,637 sgRNAs targeting 19,674 genes). The cells were treated with 10 microM DHA for 24 hours (h) and cultured in drug-free media for an additional 48h to allow outgrowth of resistant cells. After DHA treatment, BCR-ABL+ B-ALL cell viability was approximately 40% in contrast to control-treated cells that maintained >95% cell viability. After treatment, genomic DNA was isolated from the viable BCR-ABL+ B-ALL cells, amplified and subjected to Illumina sequencing to determine sgRNAs enrichment in the surviving cells.