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Abstract

Cancer Research

Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype

Published: June 17th, 2022

DOI:

10.3791/63383

1Department of Clinical Haematology, Christian Medical College, 2Department of Biotechnology, Thiruvalluvar University, 3Centre for Stem Cell Research, Christian Medical College

Abstract

Understanding clinically relevant driver mechanisms of acquired chemo-resistance is crucial for elucidating ways to circumvent resistance and improve survival in patients with acute myeloid leukemia (AML). A small fraction of leukemic cells that survive chemotherapy have a poised epigenetic state to tolerate chemotherapeutic insult. Further exposure to chemotherapy allows these drug persister cells to attain a fixed epigenetic state, which leads to altered gene expression, resulting in the proliferation of these drug-resistant populations and eventually relapse or refractory disease. Therefore, identifying epigenetic modulations that necessitate the survival of drug-resistant leukemic cells is critical. We detail a protocol to identify epigenetic modulators that mediate resistance to the nucleoside analog cytarabine (AraC) using pooled shRNA library screening in an acquired cytarabine-resistant AML cell line. The library consists of 5,485 shRNA constructs targeting 407 human epigenetic factors, which allows high-throughput epigenetic factor screening.

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Keywords Pooled ShRNA Library

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