Mutagenized cell population of 100x10e6 KBM7 cells that were selected with 100 nM YM155 over 10 days of total treatment.

We mutagenized 1x10e8 near-haploid KBM7 cells with a retroviral gene-trap vector that inserts randomly into the genome. Insertions occur genome-wide, preferentially at actively transcribed genes, disrupt the genomic locus and can result in truncation of the expressed protein product via a strong splice acceptor site. The complex mutagenized cell population was selected in the presence of 100 nM YM155 which resulted in the clonal outgrowth of resistant colonies. Drug resistant clones were pooled after 10 days of drug exposure, collected in a T175 flask and expanded to a total cell number of 3x107 cells. Genomic DNA was isolated and retroviral insertion sites were detected via an inverse PCR protocol adapted to next generation sequencing (Carette JE. et al, Nat Biotechnol 29 542-6 (2011)).