The use of modified RNAs as a therapeutic platform is an exciting new area of research made possible by recent advances in synthetic biology.
At tenOever Lab, we are developing a self-amplifying RNA that is both programmable and amenable to viral packaging. We achieve this by combining synthetic biological circuits with our self-replicating RNA platform to generate biocomputers that can perform complex operations with different sets of user-defined inputs.
The core processor for these designs are novel arthropod-specific viral RNA dependent RNA polymerase (RdRp) scaffolds which ensure both a minimalistic design and a lack of unwanted host interactions in mammalian cells. To program these constructs, we apply microRNA-enabled logic gates in conjunction with both activators and repressors of replicon biology. The idea put forth here is to generate a toolbox of adaptable and tunable logic-based replicons that will serve as a new biomaterial for genetic manipulation or gene delivery of any desired cell type or eukaryotic host.
Based on the current knowledge of these viruses, we synthesized the minimal components to enable their self-amplification as a starting platform. For V1, we generated a construct that can be in vitro transcribed and directly introduced into cells. For V2, we engineered bi-directional plasmids that generate both vRNA and mRNA from each individual plasmid. Once launched, each of these self-amplifying RNAs can be fitted with selection markers to enable functionality in non-insect hosts. Once a desired level of activity is achieved, the addition of logic gates can be applied to generate variants to define functionality as it relates to cell lineage.
Our Synthetic Biology emphasis currently has two Aims:
- Expanding on the arthropod virus-specific replicon platform
- Controlling replicon biology through miRNA-based logic gates