Date of Award

Summer 8-2018

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

Cuong Diep, Ph.D.

Second Advisor

Robert Hinrichsen, Ph.D.

Third Advisor

Robert Major, Ph.D.

Abstract

Kidney disease affects approximately 15% of the United States population, and current therapies have severe limitations. Thus, new treatment options are urgently needed. Humans cannot regenerate new kidney tissue, but zebrafish can. They do this using specialized stem cells that express the lhx1a gene. Recently, it was shown for the first time that lhx1a dimerized in a genetic assay (the yeast two-hybrid system). Deleting the LIM domain of lhx1a enhanced dimerization. Other results have also shown that deleting the LIM domain activated lhx1a, making it a constitutive transcriptional activator. Thus, it is hypothesized that lhx1a dimerization may be important for its activation. This provides a new mechanistic model for how lhx1a activity is regulated. For this reason, it is important for us to determine the dimerization domain. In this project, the CRISPR-Cas9 system is used to create a new yeast strain expressing the LexA (op)-URA3 reporter. This new strain is used for genetic screens that fail to dimerize lhx1a mutants. The results would potentially help to identify the dimerization domain. it would bring new insights into the regulation of lhx1a activity in kidney stem cells and may contribute to new regenerative therapies for kidney disease.

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