Faculty Sponsor
Wendy Olivas
Final Abstract for URS Program
Parkinson’s Disease (PD) is an age-related neurodegenerative disease. LRRK2 is one of the main PD-associated genes, as both familial mutations and its abnormal overexpression are associated with PD progression. However, underlying molecular mechanisms leading to LRRK2 abnormal overexpression are not well understood. MicroRNAs (miRNAs) are a group of small non-coding RNAs that regulate target messenger RNAs (mRNAs) by binding to a specific sequence in their 3’ UTRs (untranslated regions at the ends of mRNAs) and blocking translation of the mRNA. Previous studies have identified miR-205 as one regulator of LRRK2. However, the LRRK2 3’ UTR has many other computationally-predicted miRNA binding sites. Given that alterations in miRNA regulation are also associated with PD, the goal of this study is to examine the role of computationally-predicted miRNAs on regulation of LRRK2. The approach involves transfecting miRNA mimics into SH-SY5Y cells, which are human, dopamine-producing neuronal cells. While these cells are a great model system for PD research, they are known to be hard-to-transfect. Thus, it was important to first evaluate the efficiency of transfection of miRNAs into these cells. Transfection of a fluorescein-labeled miRNA control and a negative control miRNA into cells by electroporation was evaluated using flow cytometry and confocal microscopy. Accordingly, >80% transfection efficiency of miRNAs into our cell line was achieved, which places confidence in experiments involving miRNA transfection. Three miRNAs predicted to regulate LRRK2 (miR-130a, miR-200a and miR-185) were selected for analysis, and assays to test their regulatory effects upon transfection into SH-SY5Y cells were performed using a luciferase/LRRK2 reporter assay. miR-205 was used as the positive control and scrambled miRNA was used as the negative control. Of the miRNAs tested, miR-200a and miR130 were found to be involved in LRRK2 regulation. These results generate new knowledge on miRNA regulation of the PD-associated gene, LRRK2, which contributes to our understanding of mechanisms that may be compromised in PD.
Presentation Type
Visual Presentation
Document Type
Article