Author

Suraj Niraula

Date of Award

8-2019

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

Cuong Q. Diep

Second Advisor

Daniel Widzowski

Third Advisor

Robert Major

Abstract

Kidney disease is generally a secondary disease to other diseases like high blood pressure and diabetes. Approximately 14.8% of American adults suffer from kidney disease where dialysis and kidney transplant are the only options available for the end-stage kidney disease which are not ideal. Thus, there is an urgent need for an alternative stem cell/ regenerative therapy for treating kidney disease. Unlike humans, zebrafish can repair and regenerate their nephrons throughout their life after kidney injury and they also share significant similarities in development and composition of nephrons which makes them a promising model to study kidney development. Lhx1a is a transcriptional activator that is expressed in kidney stem cells during kidney development in zebrafish and other organisms, including mice and humans. A genetic assay showed that lhx1a dimerizes with itself and this homodimerization is supposed to activate the transcription. Deleting the LIM domain of lhx1a enhanced the dimerization and it was hypothesized that the LIM domain negatively regulates the homodimerization. Using the previously created peptide library, 20 more peptides were found to bind with lhx1a which were identified through western blot and sent for DNA sequencing. The previously found R1 peptide that binds with DNA was successfully incorporated into the RS416-intein expression plasmid and spotted with lhx1a. The R1 peptide seemed to negatively regulate the lhx1a dimerization however the higher standard deviation made the result insignificant.

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