With this project, I aim to develop methods to improve mRNA therapies by engineering structural elements into their 5’ and 3’ untranslated regions (UTRs). The primary objective is to exploit the proteomic differences between cancer cells and healthy cells so that they differentially interact with the designer UTR elements, resulting in cell-type specific expression of the mRNA. A major goal of the project is to improve the binding affinity between RNA motifs and their RNA binding proteins. This will be achieved using structural insights from cryo-EM analysis of the RNA–protein complexes and design principles from the RNA origami method. Another major goal is to use the aforementioned RNA mot...

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With this project, I aim to develop methods to improve mRNA therapies by engineering structural elements into their 5’ and 3’ untranslated regions (UTRs). The primary objective is to exploit the proteomic differences between cancer cells and healthy cells so that they differentially interact with the designer UTR elements, resulting in cell-type specific expression of the mRNA. A major goal of the project is to improve the binding affinity between RNA motifs and their RNA binding proteins. This will be achieved using structural insights from cryo-EM analysis of the RNA–protein complexes and design principles from the RNA origami method. Another major goal is to use the aforementioned RNA motifs to stabilize the mRNA in cells overexpressing the protein of interest by incorporating them as 3’UTR elements. The motifs will be unstructured in the absence of the targeted protein, resulting in rapid degradation of the mRNA, but when bound by the protein will exhibit nuclease resistance and persist for longer in the cell, achieving greater translational output. The final goal of the project is to create an mRNA that can only be translated in the presence of the RNA binding protein. This final goal will be accomplished by incorporating the RNA motifs in the 5’UTR such that they interact with the translational start sites and block translation initiation. Alternatively, cooperative binding of the RNA binding protein to the RNA motifs will cause restructuring of the motif and reveal the translation start site to the translation machinery, allowing for translation initiation. Together, these strategies will be combined to achieve translation of an exogenous mRNA in a cell type specific manner, which will allow development of mRNA therapies to selectively kill cancer cells with minimal off-target effects.

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