High-Throughput Screening of Tyrosine Kinase Inhibitor Resistant Genes in CML

Methods Mol Biol. 2016;1465:159-73. doi: 10.1007/978-1-4939-4011-0_14.

Abstract

Genome-wide RNA interference (RNAi) screening in mammalian cells has proven to be a powerful tool for identifying new genes and molecular pathways relevant to many cellular processes and diseases. For example, screening for genes that, when inactivated, lead to resistance to cancer therapeutic drugs can reveal new mechanisms for how resistance develops and identify potential targetable strategies to overcome drug resistance. Here, we describe a detailed procedure for performing a high-throughput RNAi screen using a genome-wide human short hairpin RNA (shRNA) library for identifying tyrosine kinase inhibitor (TKI)-resistance genes in a human CML cell line model.

Keywords: CML; Drug resistance; Genome-wide; Imatinib; RNAi; Tyrosine kinase inhibitor; shRNA screen.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Drug Resistance, Neoplasm*
  • Fusion Proteins, bcr-abl / genetics
  • Gene Library
  • High-Throughput Screening Assays / methods*
  • Humans
  • Imatinib Mesylate / pharmacology
  • K562 Cells
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / drug therapy
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics*
  • Protein Kinase Inhibitors / pharmacology
  • RNA, Small Interfering / genetics*
  • Sequence Analysis, DNA

Substances

  • Protein Kinase Inhibitors
  • RNA, Small Interfering
  • Imatinib Mesylate
  • Fusion Proteins, bcr-abl