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Chapters
- 00:05Title
- 02:01Construction of Lentiviral Luciferase Reporters
- 02:28Production of Lentiviral Particles: Seed Cells (Day 1)
- 03:05Transient Transfection: CaPO4/DNA Precipitation (Day 2)
- 04:23Harvest Viral Particles (Days 3-4)
- 05:02Infection of 3T3 Cells (Day 3 Onward)
- 06:12Synchronization and Change to Recording Medium
- 07:02Bioluminescence Recording of Reporter Cells
- 07:26Data Analysis and Presentation
- 08:12Results: Phase-specific Circadian Reporters
- 09:47Conclusion
Circadian clocks function within individual cells, i.e., they are cell-autonomous. Here, we describe methods for generating cell-autonomous clock models using non-invasive, luciferase-based real-time bioluminescence technology. Reporter cells provide tractable, functional model systems for studying circadian biology.
Tags
Cell-autonomous Circadian Clock RhythmsGene ExpressionLuciferase Bioluminescence ReportersMammalian Circadian ClocksSleep-wake CyclesLiver MetabolismEndogenous Circadian ClocksSuprachiasmatic Nucleus (SCN)Peripheral ClocksOscillatorsNegative Feedback MechanismCell-based In Vitro AssaysLocomotor ActivitySCN Explants Ex VivoCell-based ModelsClock FunctionTemporal ResolutionReal-time Bioluminescence RecordingFirefly Luciferase Reporter
