Document Type

Thesis

Date of Award

5-2015

School/College

College of Science, Engineering, and Technology (COSET)

Degree Name

MS in Biology

First Advisor

Assistant Professor Shodimu-Emmanuel Olufemi

Abstract

MicroRNA's (miRNAs) are short non-coding RNAs of 􁪽18-24 nucleotides. They regulate gene expression in unicellular and multicellular organisms at the posttranscription level by preventing translation and promote degradation of mRNAs. A single miRNA regulates multiple mRNAs. They govern multiple genes expression at the cellular level through cell proliferation, cell differentiation and apoptosis, and they have been implicated in many diseases such as cancer, cardiovascular, and neurodegenerative diseases. Many of the mRNAs that are targeted by miRNAs are identified by computational analysis and their functions are yet to be validated. The objective of this study is to verify the predicted mRNA targets ofhsa-miR-4259 and hsa-miR-200b; and to confirm that hsa-miR-4259 down-regulates expression ofUBF1 and NFKB2; and hsamiR- 200b down-regulates expression ofMBNL1 and MBNL3 in PC3 cells. We successfully cloned pre-hsa-miR-4259 into pmR-ZsGreen1 expression vector (i.e. GFP), creating pre-hsa-miR-4259-pmR-ZsGreen1 DNA construct, as well as overexpression of the DNA construct in PC3 cells. The result also confirmed the predicted information that 1 2 is available at the miRDB, which predicts that hsa-miR-4259 targets UBFl and NFKB2 and hsa-miR-200b targets MBNLI and MBNL 3 to downregulate their proteins expression in PC-3 cells.

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