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合并的shRNA文库筛选,以确定调节耐药性表型的因素
JoVE Journal
Cancer Research
Author Produced
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JoVE Journal Cancer Research
Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype
DOI:

14:51 min

June 17, 2022

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Chapters

  • 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

Automatic Translation

使用慢病毒 shRNA 池的高通量 RNA 干扰 (RNAi) 筛选可以成为检测恶性肿瘤中治疗相关合成致死靶标的工具。我们提供一种合并的shRNA筛选方法来研究急性髓系白血病(AML)中的表观遗传效应子。

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