Biological Systems Engineering, Department of
Date of this Version
Fall 12-4-2015
Document Type
Article
Citation
Nguyen AL. High Throughput Screening of Priming Candidates for Impact on Nonviral Gene Delivery. University of Nebraska-Lincoln. 2015.
Abstract
Priming, in the context of nonviral gene delivery, is the treatment of cells with a compound prior to gene transfer that enhances transfection efficiency and/or transgene expression. Essentially, it is the application of an adjuvant approach to gene delivery. Effective transfection strategies may require priming to compete with the efficiency of viral transduction in order to achieve clinically relevant efficiency and expression in vivo. To search for priming compounds, a high throughput screen of the NIH Clinical Collection was performed using 25kDa b-PEI, an EGFP/luciferase plasmid, and HEK293T cells. The EGFP reporter was multiplexed with Hoechst 33342 and Resazurin fluorescence to measure transfection efficiency, transgene expression, proliferation, and viability of the cells in response to priming with the screened NCC compounds and PEI transfection. The screen identified dozens of compounds and several compound classes that appear to affect transfection through modulation of mitochondrial dysfunction in response to the toxicity of PEI complexes. With further investigation and development, the mechanisms by which these and other priming compounds are affecting gene transfer can be understood and applied towards the development of efficient gene delivery strategies.
Advisor: Angela K. Pannier
Comments
A THESIS Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Master of Science, Major: Agricultural and Biological Systems Engineering, Under the Supervision of Professor Angela K. Pannier. Lincoln, Nebraska: November 2015
Copyright 2015 Albert Nguyen