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Capitoli
- 00:05Titolo
- 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
Circadiane klokken functioneren binnen de individuele cellen, dat wil zeggen, ze zijn cel-autonoom. We beschrijven werkwijzen voor het genereren van cel-autonome klok modellen niet-invasieve, luciferase-gebaseerde real-time bioluminescentie technologie. Reporter cellen handelbaar, functionele modelsystemen voor het bestuderen van circadiane biologie.
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
