Year: 2015
Type: article
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
Source: Lincoln (University of Nebraska)
Author: Albert Nguyen
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2190275523