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En Multiplexed Luciferase-baseret Screening Platform for forhører Kræft-associeret signaltransduktion i dyrkede celler
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JoVE Journal Medicine
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

En Multiplexed Luciferase-baseret Screening Platform for forhører Kræft-associeret signaltransduktion i dyrkede celler

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

Opnå et system level forståelse af cellulære processer er et mål for moderne cellebiologi. Vi beskriver her strategier for multiplexing luciferase journalister af forskellige cellulære funktion endepunkter at afhøre genfunktion hjælp genom-skala RNAi biblioteker.

Tags

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

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