SEM cell line, reference library, replicate 2
A sub-genomic CRISPR/Cas9 screen was designed to target 36 RNA binding protein genes , 12 positive control genes and 28 Non-targeting guide RNAs. For each gene, 5 guide RNAs were designed using sgRNA designing tools from the Broad Institute. Pooled sgRNA were then cloned into pLKO5.sgRNA.EFS.tRFP lentiviral vector using standard protocols. Next, B-ALL cell lines with MLL-AF4 translocation (SEM cells) and without MLL-AF4 translocation (NALM-6) were  stably transduced with pLentiCas9-GFP20  lentiviral preparation and Cas9 expression in these cells checked over the time prior to CRISPR screen. Next, lentiviral pools were made from pLKO5.sgRNA.EFS.tRFP vector, and titers were calculated from SEM and NALM-6 cell transduction