title: Transcriptomic analysis of imatinib-resistant K562 cells with knocked down Pleomorphic Adenoma Gene 1 (PLAG1) to identify the targets
design: Transcriptomic analysis of imatinib-resistant K562 cells with knocked down Pleomorphic Adenoma Gene 1 (PLAG1) to identify the targets
library_construction_protocol: Imatinib resistant K562 cells, which were established in lab, were transduced with to lentiviral particles containing shRNA 2655.2 to knockdown PLAG1 (from Sigma), and lentiviral particles containing pLKO.1-empty vectors were generated in the lab. Puromycine based selection was carried out and polyclonal populations were established for empty vector  and PLAG1 knockdown. Further, with flow assisted cell sorting monoclonal populations were generated and expanded. RNA was extracted from both empty vector containing cells and PLAG1 knockdown cells. RNA was extracted from imatinib-sensitive and -resistant K562 cells, using AllPrep DNA/RNA/miRNA Universal Kit (Qiagen, MD, USA: catalogue no. 80224) according to the manufacturer's recommendations. RNA samples were subjected to the library preparation as per NEB Ultra RNA-Seq Library Prep kit protocol (Cat# E7530L).