The examination of bacteriophage M13-based miniphagemids as a platform for the delivery of genetic material for neuronal applications

Abstract

M13-derived vectors have been shown to deliver foreign genetic material in vitro as well as bypass the blood-brain barrier, offering a promising approach to gene therapy applications that target the brain. Miniphagemid particles combine the capsid architecture of M13, which enables cellular targeting through peptide display, with a minimized genetic sequence, creating a highly modular vector. Despite the potential of these particles, their novelty and unique biology have limited the examination of their interaction with neuronal cells. This thesis examines the tailoring of miniphagemid particles to neuronal applications though neuropeptide display and transcriptional control. The outlined changes did not result in a significant increase in transfection efficiency in neuronal cells, instead revealing the unsuitability of the promoter used and ssDNA. Although, the specific application to neuronal cultures was unsuccessful, the use of a novel characterization method in miniphagemid particle analysis, specifically multi-angle dynamic light scattering, illustrated a substantial issue in current production methods; highlighting the current overreliance on functional quantification methods and supporting multi-angle dynamic light scattering as a physical method to determine miniphagemid particle purity.