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Genome-wide CRISPR screen in SUM149 cells
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SUM149-Cas9 cells were generated by transduction of SUM149 cells with a Cas9-bsd lentivirus and selection in 7 µg/ml blasticidin. Cells were infected at multiplicity of infection (MOI) 0.3 with a previously published genome-wide human lentiviral CRISPR library68. Cells were selected with puromycin and then placed under EX527 selection at a concentration that killed all non-infected SUM149 cells (100 μM). sgRNA sequences from resistant cells were PCR amplified and sequenced.
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