Date of Award

4-2023

Degree Type

Thesis

Degree Name

Master of Science

Department

Biology

Program

Biology (MS)

First Advisor/Chairperson

Dr. Robert Belton

Abstract

10-15% of couples worldwide are affected by infertility. The leading cause of infertility is implantation failure. Molecular communication in precise time and space between an implanting embryo and a receptive uterus is essential for successful implantation. Basigin has been shown to have vital functions within the implantation process. Basigin is best known for inducing the expression of matrix metalloproteinases. Basigin has been shown to induce MMP expression in vitro in human endometrial stromal cell lines (HESC). Understanding how basigin induces matrix metalloproteinases within normal cellular conditions will help researchers understand how these signaling cascades flow in the framework of implantation. In order to do this, I attempted to characterize a basigin knock-out cell line using CRIPSR/cas9 in human endometrial stromal cells. Sixteen potential candidate clones were screened for the knock-out using PCR and western blotting. I determined that the donor plasmid was inserted into the genomes, but not in the intended location. Upon western blotting, all clones contained basigin protein expression. This could be due to basigin being a key gene in cell regulatory processes. Soluble basigin was then used to stimulate wildtype HESC lines for 0, 6, or 24 hours to characterize MMP protein expression and activation using immunoblotting and zymography. I found the highest expression fold changes in MMP-1, -2, and -9 at 6 hours, and MMP-2 at 24 hours. Zymography showed no difference in time. Future work is recommended to pinpoint peak protein expression for MMPs in HESCs following soluble basigin stimulation.

Access Type

Open Access

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