Dasatinib Resistant K562 cell lines
A project to explore the acquisition of drug resistance mechanisms in a K562 erythroblastic leukemia cell line, with induced resistance to the tyrosine kinase inhibitor, dasatinib. Cells were exposed to increasing concentrations of dasatinib over ~11 months, up to a maximum dose of 200 nM dasatinib. Sequencing was performed in a selection of dasatinib dose escalation intermediates, ranging from 0 nM to 200 nM dsasatinib.Sequencing libraries were prepared using the NEXTFlex Rapid Directional RNA Seq Library Prep Kit (Perkin Elmer, Inc., Waltham, MA), depleting ribosomal and non-coding RNA. Quality control of RNA was performed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA), using the Bioanalyzer RNA 6000 Nano assay. Libraries were sequenced on the Illumina MiSeq instrument (Illumina Inc., San Diego, CA), using a 150 cycle to produce 2x75 bp paired end reads. Sequencing data was uploaded to the Illumina BaseSpace Sequencing Hub (Illumina Inc.), which was used for FastQC quality control.Results were used for differential gene expression analysis (EdgeR), screening for SNV and indel mutations (GATK) and analysis of gene fusions (FusionCatcher).