Development of Novel Circadian Reporter

Loading...
Thumbnail Image

Persistent link to this item

Statistics
View Statistics

Journal Title

Journal ISSN

Volume Title

Title

Development of Novel Circadian Reporter

Published Date

2022-08

Publisher

Type

Thesis or Dissertation

Abstract

Adaptations to light and dark cycles on Earth are integral to the survival of all organisms. Indeed, all living organisms retain some form of biological rhythms that ebb and flow with daily environmental stimuli. Importantly, disruptions to circadian rhythms (CRs) are associated with reduction in life/health span and numerous metabolic disorders. As such, there is substantial interest in regulating CRs to improve health outcomes. In mammals, these CRs are generated by a transcriptional-translational feedback loop composed of core clock proteins BMAL1, CLOCK, PERIOD and CRY. Other proteins include RORa and REV-ERBa, which regulate expression of BMAL1 and CLOCK. Alongside light stimuli, CRs are sensitive to environmental cues such as food intake. There has been recent interest in the role that presence of key nutrients, specifically lipid species, plays in regulating CRs. However, the mechanism by which these metabolites act upon the circadian clock is unknown. One limitation of CR-centered research is that it is inherently low-throughput. CR studies typically incur a large financial cost and require large amounts of labor to process and analyze biological samples due to the sheer number of time points and biological replicates. As such, the field would benefit from an accurate circadian reporter that is sensitive to changes to clock gene expression. Here, we present a novel circadian reporter that can streamline circadian rhythm data collection and analysis with the use of PERIOD2-promoter driven fluorescent reporter. We tested the efficacy of the reporter in AML12 cells stably expressing it and treated them with several known circadian inducers to determine complete and robust circadian profiles. In future directions, we aim to optimize experimental design parameters to enhance performance of the novel circadian reporter.

Keywords

Description

University of Minnesota M.S. thesis.August2022. Major: Biochemistry, Molecular Bio, and Biophysics. Advisor: Douglas Mashek. 1 computer file (PDF); viii, 46 pages.

Related to

Replaces

License

Series/Report Number

Funding information

Isbn identifier

Doi identifier

Previously Published Citation

Suggested citation

Alvarez, Jonas. (2022). Development of Novel Circadian Reporter. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/243058.

Content distributed via the University Digital Conservancy may be subject to additional license and use restrictions applied by the depositor. By using these files, users agree to the Terms of Use. Materials in the UDC may contain content that is disturbing and/or harmful. For more information, please see our statement on harmful content in digital repositories.