The Effect of an miRNA Sponge on miR-124 Target Tumor Suppressors

Undergraduates: Karthika Kandala, John Noto, Casey Schmidt

Faculty Advisor: Greg Matera
Department: Biology

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MicroRNAs (miRNAs), a class of noncoding RNAs, have been examined as potential regulators of cancer cell progression and metastasis pathways. miRNAs bind to mRNA transcripts and target them for degradation, thereby, serving to control gene expression. In this project, I used tRNAs to generate tRNA intronic circular (tric)RNAs to make an exogenously expressed miRNA sponge. miR-124 has been known to reduce the proliferation of the breast cancer cell, MCF-7. It is also the most abundantly expressed miRNA in neuronal cells. A miR-124 sponge construct that contains four binding sites for miR-124 was designed using a bioinformatic software (ApE). The miR-124 sponge was transfected into HEK293T cells and the RT-PCR assay showed that the sponge was being expressed. The next step would be to conduct a cell growth assay to assess the growth profiles of miR-124 inhibited cells, which will be used to look at the gene expression of miR-124 targets using RT-PCR and Western Blotting.