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Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype
JoVE Journal
Krebsforschung
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JoVE Journal Krebsforschung
Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype
DOI:

14:51 min

June 17, 2022

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Kapitel

  • 00:00Introduction
  • 00:24Selection of the Most Potent Promoter to Obtain Persistent and Prolonged Expression of the shRNAs
  • 02:01Preparation of Pooled Lentiviral Human Epigenetic Factor shRNA Library
  • 04:48Estimation of the Transduction Efficiency of Lentiviruses
  • 06:03Transduction of Pooled Epigenetic shRNA Library in the Drug-Resistant Cell Line
  • 07:20Enrichment of GFP Positive Cells
  • 07:54Dropout Screening to Identify Epigenetic Factors Mediating Drug Resistance
  • 09:20Amplification of the Integrated shRNAs by PCR
  • 11:08Next-Generation Sequencing and Data Analysis
  • 11:57Representative Results
  • 14:30Conclusion

Summary

Automatische Übersetzung

High-throughput RNA interference (RNAi) screening using a pool of lentiviral shRNAs can be a tool to detect therapeutically relevant synthetic lethal targets in malignancies. We provide a pooled shRNA screening approach to investigate the epigenetic effectors in acute myeloid leukemia (AML).

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