Journal
/
/
Cribado agrupado de la biblioteca de shRNA para identificar los factores que modulan un fenotipo de resistencia a los medicamentos
JoVE Journal
Cancer Research
Author Produced
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Cancer Research
Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype
DOI:

14:51 min

June 17, 2022

, , , , ,

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

El cribado de interferencia de ARN (ARNi) de alto rendimiento utilizando un grupo de shRNAs lentivirales puede ser una herramienta para detectar dianas letales sintéticas terapéuticamente relevantes en neoplasias malignas. Proporcionamos un enfoque de detección de ARNt agrupado para investigar los efectores epigenéticos en la leucemia mieloide aguda (LMA).

Related Videos

Read Article