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miRNAs represent a diverse family of eukaryotic biological regulators, which target specific mRNA sequences to prevent translation. Diseases such as Alzheimer’s, cancer, and diabetes exhibit aberrant gene expression. Targeting this via RNA interference has the advantage of being safer than conventional gene therapeutic methods due to lack of integration into the host genome. This motivates our novel approach: a modular, self-replicating RNA system. Specifically, we will create an independently replicating RNA operon in human (HeLa) cells using a HCV derived RNA-dependent RNA polymerase (RdRp, RNA replicase). To test this system, we will produce miRNA-29a to downregulate the dipeptidyl peptidase-IV (DPP-IV), which is elevated in Type 2 diabetes and is the target of many drug studies. Additionally, we will incorporate a control module in the form of a multi-input sensing feedback mechanism which will function as a logic circuit to regulate expression of miRNA.
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