Poster Session 3, 1:15 PM - 2:00 PM: Campus Center Auditorium [A33]

Investigating the Repressor Sequestration Model of Cilia Length Regulation

Presenter: Emily Hope Wassouf

Faculty Sponsor: Jason M. Brown

School: Salem State University

Research Area: Biology

ABSTRACT

Cilia are hairlike projections on the surface of many cell types that provide the cell with functions such as the ability to swim and sense the external environment (Brown and Witman 2014). Defects in cilia can cause diseases in humans, or ciliopathies. While many genes responsible for building cilia are identified, the complete mechanisms regulating these genes remain unknown. The Repressor Sequestration Model, introduced in Perlaza et al. (2022), provides a possible method of cilia gene regulation. This model proposes the presence of a repressor that, in cells with steady-state cilia, remains concentrated in the cilia, cell body, and nucleus, where it blocks cilia gene upregulation. According to the model, upon deciliation and immediate regrowth, the repressor is sequestered into cilia and away from the nucleus, allowing cilia building genes to become derepressed and their expression levels to increase. By using the biciliated green alga, Chlamydomonas reinhardtii, we developed a workflow for generating mutants with a disrupted repressor gene. Random insertional mutations in Chlamydomonas were generated via electroporation and the resulting transformants grown. Mutant colonies were individually picked, then screened via biochemical and microscopic analysis. Mutants with altered levels of cilia gene expression could suggest a disrupted repressor gene. Currently, screenings are still being done with the hope of finding a mutant with constitutively high levels of cilia gene expression. After identifying said mutant, we will use RESDA-PCR (González-Ballester et al. 2005) to determine the location of the mutation, and the possible identity of the gene or genes it is impacting. 

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