Imani Bethel

Document Type

Thesis

Date of Award

5-19-2014

School/College

College of Science, Engineering, and Technology (COSET)

Degree Name

MS in Biology

First Advisor

Shodimu-Emmanuel Olufemi

Abstract

Alternative pre-rnRNA splicing contributes to protein diversity and protein expression of aberrantly spliced mRNA contributes to human diseases. Many splicing regulators have been identified and studied, but RNAs that regulate these splicing regulators have not been fully elucidated. One of the RNAs that regulates the splicing regulators is the non-coding RNA, the homo sapien microRNA (hsa-miR-130b). To understand the function(s) and effects of non-coding RNAs on aberrant mRNA splicing regulators, we identified in the literature a gene, Gemin5, an important component pre mRNA splicing regulators. We searched the miR Base database and identified a microRNA (miRNA), hsa-miR-130b, that target mRNA of Gemin5, which is one of several splicing factor families. The Gem-associated protein (Gemin) family and the Muscle blind-like (MBNL) family are both involved in splicing. Gemin5 is a component 1 in alternative splicing of pre-mRNA. Disturbance of the spliceosome complex would result in aberrant splicing of pre-mRNA; therefore, the objective of this study is to determine if hsa-miR-130b actually target mRNA of Gemin5 to downregulate protein expression of Gemin5 in PC-3 cancer cells. We hypothesize that overexpression of hsa miR-130b in PC-3 cells will target mRNA of Gemin5 and Muscle blind-like 1 (MBNL1) to downregulate protein expression of Gemin5 and MBNL1. For the first time, we show that overexpression of hsa-miR-130b downregulates protein expression of Gemin5 and MBNL1, which are members of the pre-mRNA splicing factors. Our results confirm that hsa-miR-130b does indeed target Gemin5 and MBNL1 mRNAs in PC-3 cells

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