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Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter
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Cancer Research
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JoVE Journal Cancer Research
Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter

Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter

DOI:
10.3791/60582-v

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11:32 min

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March 27, 2020

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,
1Department of Molecular & Cellular Physiology,Albany Medical College

Chapters

  • 00:04Introduction
  • 00:39Lentiviral Vector Library Preparation
  • 01:24Packaging of the Arrayed Lentiviral Library
  • 03:35Cell Infection for Screening
  • 04:48Cell Seeding for Transfection of Dual-Luciferase Reporter
  • 06:27Dual-Luciferase Reporter Transfection
  • 07:35Dual-Luciferase Activity Quantification
  • 09:02Results: Representative Arrayed Library Screening
  • 10:47Conclusion

Summary

Automatic Translation

Automatic Translation

To identify novel regulators of transcription factors, we developed an approach to screen arrayed lentiviral or retroviral RNAi libraries using a dual-luciferase-based transcriptional reporter assay. This approach offers a quick and relatively inexpensive way to screen hundreds of candidates in a single experiment.

Tags

Keywords: Transcription FactorMedium-throughput ScreeningArrayed LibrariesDual-luciferase ReporterLentiviral VectorRetroviral AssayBacterial Miniprep293FT CellsTransfectionPsPAX2VSVGCell Culture

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