Applied Magnetic Forces Enhance Nanoparticle Based Gene Delivery and Characterize Intracellular Rheology and Transport

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Ford, Kris Iniko
- Affiliation: School of Medicine, UNC/NCSU Joint Department of Biomedical Engineering

Abstract

Magnetically applied forces present the ability to manipulate paramagnetic nanoparticles for a wide variety of biomedical applications. In vivo gene delivery offers the promise of new ways to treat disease at a genetic level, but it unfortunately suffers from low efficiencies. Nanoparticles present a platform for non-viral gene therapy but delivery must be optimized to make them a viable option for treatment. This work describes how magnetic forces can be applied to paramagnetic nanoparticles to enhance the delivery of antisense oligonucleotides for correction of mRNA splicing errors. Applied oscillating forces can be used to stimulate certain endocytic pathways, which acts to more than double transfection efficiency. Once a nanoparticle enters the cell, it is surrounded by the complex intracellular environment. To understand the environment that the nanoparticle enters, intracellular rheology is probed with paramagnetic microbeads. A protocol for introducing 1 micron microbeads into cells is presented. By manipulation with 3-dimensional force microscopy, the mechanical features of the cell can be characterized. Additionally, the in vivo transport due to molecular motors is also characterized. Stall force on an intracellular 1 micron bead, caused by dynein motors, is determined. This represents the force exerted on a vesicle by the motors, as it is being transported towards the nucleus, the target of the oligonucleotide delivery.

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