THE DESIGN, GENERATION, AND INVESTIGATION OF DUMBBELL-SHAPED MINIMAL DNA VECTORS FOR DELIVERY OF CODING AND NON-CODING RNA

Abstract

Dumbbell-shaped DNA (Db) vectors are a new generation of recombinant DNA vectors, investigated for the delivery of functional non-coding or coding RNA. By virtue of their design, they have marked benefits over existing state-of-the-art vector systems. This thesis contributes towards improving three facets of Db vector technology: (a) Production: A novel PCR-based strategy is described for the production of Db vectors expressing shRNA; (b) Delivery: Novel peptides are investigated for their cell penetrative properties and their propensity to aide in cellular delivery of appended Db vectors expressing shRNA; and (c) Application: Db vector technology is investigated for the delivery of components of the RNA-guided CRISPR-Cas9 DNA editing system.