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약물 내성 표현형을 조절하는 인자를 식별하기 위한 풀링된 shRNA 라이브러리 스크리닝
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

렌티바이러스 shRNA의 풀을 이용한 고처리량 RNA 간섭(RNAi) 스크리닝은 악성종양에서 치료적으로 관련된 합성 치사적 표적을 검출하는 도구가 될 수 있다. 우리는 급성 골수성 백혈병 (AML)에서 후성 유전 학적 이펙터를 조사하기 위해 풀링 된 shRNA 스크리닝 접근법을 제공합니다.

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