Researcher(s)
- Robyn Logue, Engineering Undecided, University of Delaware
Faculty Mentor(s)
- Kevin Solomon, Department of Chemical and Bimolecular Engineering, University of Delaware
- Akash Vaidya, Department of Chemical and Bimolecular Engineering, University of Delaware
Abstract
RNA has immense potential in vaccines and medicine but requires nanoparticle carriers for protection and delivery. Rod-shaped plant viruses are protein assemblies that naturally coat, protect, and deliver single-stranded RNA genomes, thus representing a powerful platform for RNA delivery applications. Their viral coat proteins can also assemble around arbitrary RNA templates containing an origin of assembly sequence (OAS) to form non-infectious, virus-like particles (VLPs). Our group recently produced Barley Stripe Mosaic Virus (BSMV) VLPs in bacteria, enabling flexible engineering of coat proteins for targeted biological functionality. However, bacterial BSMV VLP RNA templates lack eukaryotic elements needed for RNA vaccine applications. We resolved this issue through cell-free (in vitro) assembly of BSMV VLPs. We developed a workflow for disrupting VLPs, removing their RNA, and reassembling the coat protein onto synthetic RNA templates. In vitro RNA can be precisely designed to have targeted length, sequence, and eukaryotic elements such as 5’ caps and 3’ poly(A) tails. Furthermore, in vitro RNA production allows the incorporation of modified bases for tailored immune responses and translation efficiencies. Assembling BSMV VLPs in vitro will further their potential as RNA vaccine carriers.