In Vivo Gene Delivery with Pulsed Electrical Fields.

R. W. MALONE. Gene Delivery Alliance, Inc., Jefferson, MD 21755. Email: gtinventor@hotmail.com

This looks kind of interesting, considering…

It’s from the 1990s, likely 1994.

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Non-viral gene delivery research currently confronts three significant obstacles, each of which contribute to the modest and transient levels of transgene expression that are often observed; 1) Exogenous polynucleotides are rapidly degraded by cytoplasmic and extracellular nucleases, 2) when used in vivo, most non-viral gene delivery technologies are very inefficient for delivering the payload into cytoplasm, and 3) peak trans-gene expression is typically observed in the first few days and then rapidly declines (often to undetectable levels). Results obtained from studies addressing each of these issues will be reviewed, with a particular focus on unpublished work involving electroporative lung transfection and transplantation in the rat model. The results to be presented demonstrate that DNA degradation and clearance may be reduced by formulation with a broad-spectrum nuclease inhibitor, electroporation may be safely used in vivo to efficiently permeabilize cells enabling plasmid uptake directly into cytoplasm, and an efficient housekeeping promoter known to provide prolonged transgene expression in lung can yield sustained transgene expression after electroporative delivery. Lung electroporation conditions were optimized at four 300V/cm, 20ms pulses administered at 1Hz by flat electrode plates. After transplanting lungs transfected with a human ubiquitin promoter driving luciferase expression, at least 40 days of high- level transgene expression were observed. By combining these factors with optimized DNA/ATA formulations, the data indicates that levels of transgene expression in transfected lung tissue may be increased by 15,000 fold relative to ‘‘naked DNA’’

hmmmm

This photo is not from 1994

A horse with Robert Malone but not in 1994