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In Vitro Bioluminescence Assay to Characterize Circadian Rhythm in Mammary Epithelial Cells
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Journal JoVE Génétique
In Vitro Bioluminescence Assay to Characterize Circadian Rhythm in Mammary Epithelial Cells

In Vitro Bioluminescence Assay to Characterize Circadian Rhythm in Mammary Epithelial Cells

DOI:
10.3791/55832-v

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11:56 min

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September 28, 2017

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, , , ,
1Department of Environmental and Occupational Health, School of Public Health, NIEHS Center for Environmental Exposures and Disease, Environmental and Occupational Health Sciences Institute,Rutgers University, 2Veterinary Drugs & Biologics Division,Animal and Plant Quarantine Agency

Chapitres

  • 00:05Titre
  • 00:53Concstruction of PER2 Promoter Driven Destabilized Luciferase Reporter Vector
  • 03:06Transforming Ligated Vector pGL[hPer2P/Luc2P/Neo] to E. coli
  • 05:28Transient Transfection of Cells with the Circadian Vector
  • 07:46Establishment of the In Vitro Bioluminescence Assay in Transiently Transfected MCF10A Cells
  • 08:52Results: Effects of Chemicls on the Cellular Biological Clock
  • 10:16Conclusion

Summary

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An in vitro bioluminescence assay to determine cellular circadian rhythm in mammary epithelial cells is presented. This method utilizes mammalian cell reporter plasmids expressing destabilized luciferase under the control of the PERIOD 2 gene promoter. It can be adapted to other cell types to evaluate organ-specific effects on circadian rhythm.

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