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A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
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JoVE Journal Medicine
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

DOI:
10.3791/50369-v

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10:13 min

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July 03, 2013

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1Department of Cell Biology,UT Southwestern Medical Center

Chapters

  • 00:05Title
  • 01:54Preparation of Cells for siRNA and Reporter Transfection
  • 03:19siRNA and Reporter Transfection of Cells
  • 03:22Preparing Reporter Construct DNA
  • 04:10Preparing siRNA/DNA Transfection Mixtures
  • 05:42Adding Transfection Mixtures to Cells
  • 06:21Determine Luciferase Activities from Cell Samples
  • 08:04Results: High-throughput Interrogation of Gene Function in Wnt/β-catenin, p53, and Kras Singaling
  • 09:53Conclusion

Summary

Automatic Translation

Automatic Translation

Achieving a systems level understanding of cellular processes is a goal of modern-day cell biology. We describe here strategies for multiplexing luciferase reporters of various cellular function end-points to interrogate gene function using genome-scale RNAi libraries.

Tags

Keywords: Multiplexed Reporter SystemRNA InterferenceRNAiGenome-scale Gene FunctionCancer Cell VulnerabilitiesHigh-content ScreeningMolecular Targeted Therapy

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