Identification of miR-29a targets in mitochondrial DNA

Zeynep Ozturk

Studenteropgave: Semesterprojekt

Abstrakt

MiRNAs are post-transcriptional regulators of protein expression and they bind to complementary sequences in the 3’UTRs of their target mRNA either performing transcript degradation or translational inhibition. It is accepted that they play a role in the defect of β-cells to secrete enough insulin so trigger type 2 diabetes mellitus (T2DM). T2DM is a common, complex, metabolic disorder. It can characterize with these defects; insulin resistance in peripheral tissues and beta‐cell dysfunction or reduced beta cell mass. Due to these defects blood glucose level increase. At the same time there is a real that T2DM is not because of only these reasons. Also genetic actions seems to be involved in the development of T2DM. This project focuses on a microRNA that has been shown to be involved in glucose‐induced beta‐cell dysfunction. The key findings from other studies were that miR‐29a is up‐regulated by glucose in beta‐cells and decrease glucose‐stimulated insulin secretion by inducing decreased activity of the electron transport chain. Recently two publications have shown that microRNAs are present in the mitochondria, suggesting that microRNAs can regulate mitochondrial gene-expression. So in this project the target sequence was searched for miR29-a. Based on this information it was tried to identify any targets of miR-29a in the mitochondrial encoded subunits of the electron transport chain. In this project we identified potential targets of miR-29a in: in ND6, ND5 and D-LOOP. The oligos were designed as having a predicted target sequence for miR29-a and transferred to a vector which has luc2 gene. These vectors were copied in E.coli cells then transfected HEK cells for luciferase assay. Unfortunetly the luciferase assay’s data and t-test showed that there is no target sequence in ND6. During the experiments because of the unknown reasons we could not use other oligos ND5 and D-LOOP. But for say a certain answer this experiment must repeat.

UddannelserMolekylærbiologi, (Bachelor/kandidatuddannelse) Bachelor el. kandidat
SprogEngelsk
Udgivelsesdato8 jun. 2012

Emneord

  • ND6
  • type 2 diabetes
  • mitochondria
  • microRNA
  • diabetes
  • miR-29a