A Lentiviral Fluorescent Reporter to Study Circadian Rhythms in Single Cells

Area of Science:

• Molecular Biology
• Chronobiology
• Cell Biology

Background:

• Circadian rhythms are ~24-hour oscillations driven by cell-autonomous molecular clocks.
• These rhythms regulate cellular responses to external signals, impacting differentiation and apoptosis.
• Existing methods for single-cell circadian visualization are limited by genomic engineering and clonal expansion.

Purpose of the Study:

• To develop novel, accessible tools for visualizing and quantifying single-cell circadian dynamics.
• To enable the study of circadian rhythms in diverse cell types without complex genetic manipulation.

Main Methods:

• Development of fluorescent circadian reporters utilizing the murine REVERBα/Nr1d1 gene.
• Delivery of reporters via lentiviral transduction.
• Compatibility with time-lapse single-cell microscopy for dynamic observation.

Main Results:

• Reporters generate oscillatory signals dependent on a functional molecular clock.
• Circadian dynamics parameters, such as phase, can be determined at the single-cell level.
• Simple and efficient delivery facilitates application across a wide variety of cell types.

Conclusions:

• The novel fluorescent reporters offer a broadly applicable method for studying single-cell circadian dynamics.
• These tools expand research opportunities into the impact of circadian rhythms across biological systems.
• Facilitates deeper understanding of cellular decision-making influenced by circadian timing.