AbstractsBiology & Animal Science

The creation of a cell line stably over-expressing miR-29a

by Anne Mette Larsen




Institution: Roskilde University
Department:
Year: 2015
Keywords: microRNA; Diabetes
Record ID: 1122083
Full text PDF: http://rudar.ruc.dk/handle/1800/20392


Abstract

Diabetes Mellitus (diabetes) is today a worldwide problem, accounting for approximately 1.5 million deaths worldwide in 2012. No cure is currently available in treating diabetes hence; there is a need for better early diagnostic biomarkers and therapeutics. One potential molecule could be micro ribo-nucleic acid (miRNA or miR). It has for example been shown that hsa-miR-29a is up-regulated in β-cells, when glucose levels are elevated. The muscles are the greatest consumer of glucose in the body, and therefore it could be of interest to investigate the expression of miR-29a in this kind of tissue, and furthermore miR-29a’s role in the regulation of the metabolism. One way to examine this is to generate a muscle cell line, which would stably overexpress the miR-29a. Different methods have, in this study been used in doing so. First, a cloning was executed making a valid control plasmid. Then a transfection, infection and selection were performed, creating the C2C12 cell line that should stably overexpress miR-29a. This creation was however not completely successful, but knowledge concerning the infection procedure has been gained. This knowledge could be used in future studies to investigate miR-29a’s role in muscle cells in regards to diabetes.