Collaborative: Improved Synthetic Vectors by Electrostatic Co-Extrusion

University Of Colorado At Denver, Aurora CO

Investigators

Abstract

Lengsfeld 0433811 This proposal was an unsolicited submission. This proposal aims to develop a new methodology to prepare gene therapy vectors by an electrostatic co-extrusion (ECE) process. This process would make it possible to encapsulate DNA using charge-neutral bilayers, potentially reducing the cytotoxicity of the vectors and improving the bioavailability of the DNA by not having a strong attachment of lipid to the DNA. Importantly, the encapsulation would be performed with very little loss of DNA, giving ECE a major advantage over pressure extrusion or other liposome preparation methods. The specific aims of this research are to: (1) demonstrate the ability of electrostatic co-extrusion to efficiently encapsulate naked plasmid DNA, DNA condensates, and siRNA within bilayers of varied lipid composition, (2) characterize vectors created by electrostatic co-extrusion to classical techniques through the assessment of homogeneity and range in size, zeta potential, and DNA content, (3) quantify and optimize DNase resistance, serum stability, and in vitro transfection rates of vectors produced by electrostatic co-extrusion, and (4) investigate approaches that would allow this technique to be scaled up for bulk manufacturing.