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Updated: Sep 2, 2026
Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia
Published on: September 18, 2013
High-Throughput Screening of Tyrosine Kinase Inhibitor Resistant Genes in CML
Leyuan Ma1,2, Justine Roderick3, Michelle A Kelliher3
1Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, 364 Plantation Street, Worcester, MA, 01605, USA. leyuan.ma@umassmed.edu.
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.
Insights
Genome-wide RNA interference screens identify genes involved in drug resistance. This study details a high-throughput method using short hairpin RNA libraries to find genes conferring resistance to tyrosine kinase inhibitors in chronic myeloid leukemia cells.
Area of Science:
- Genetics and Genomics
- Molecular Biology
- Cancer Research
Background:
- Genome-wide RNA interference (RNAi) screening is vital for discovering genes and pathways in cellular processes and diseases.
- Identifying genes conferring resistance to cancer therapeutics can reveal resistance mechanisms and suggest strategies to overcome it.
Purpose of the Study:
- To describe a high-throughput RNAi screening procedure.
- To identify genes associated with tyrosine kinase inhibitor (TKI) resistance in a human chronic myeloid leukemia (CML) cell line model.
Main Methods:
- Utilizing a genome-wide human short hairpin RNA (shRNA) library.
- Performing high-throughput RNAi screening in a human CML cell line.
- Analyzing gene inactivation effects on TKI resistance.
Main Results:
- The study provides a detailed protocol for conducting genome-wide shRNA screens.
- The method successfully identifies genes involved in TKI resistance.
Conclusions:
- High-throughput RNAi screening is an effective approach for identifying drug resistance genes.
- This methodology can advance the understanding of cancer therapeutic resistance and inform new treatment strategies.
