Date of Award

6-2013

Document Type

Open Access

Degree Name

Bachelor of Science

Department

Biology

First Advisor

Brian Cohen

Language

English

Keywords

birth control, contraception, proteins, hormone

Abstract

Oral contraceptive pills use high doses of hormone to suppress the reproductive arm of the endocrine system in women. However, these drugs can have harmful long term side effects such as increased risk of stroke, heart attack, and breast cancer. Therefore, it has become an important public health goal to look for alternative ways of providing contraception. Aptamers, oligonucleotides that bind to specific molecules due to their unique 3-dimensional structure, are one alternative that has yet to be explored. The goal of our research is to develop an RNA aptamer that will bind selectively to the hFSHR. We first incubated a large pool containing random sequence oligonucleotides with wild type HEK293 cells (which do not have hFSHR) as a counter-selection. Many of the RNA molecules bound to the protein receptors on the cell surface. The unbound molecules were then incubated with HEK293 cells expressing the hFSHR. The oligonucleotides that bound to the hFSHR were eluted, isolated, and then re-amplified. The process of selection was repeated while increasing stringency of binding each time. We were able to successfully perform SELEX on our RNA pool at 4°C and 22°C. We also conducted nanodrop experiments to confirm that our oligonucleotides are indeed binding specifically to the hFSHR. We hope to continue refining our aptamer at higher temperatures such as 30°C and 37°C, determine the exact nature of the oligonucleotide-protein binding complex, and eventually begin in vivo trials to determine efficacy.

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