Electroporation of the Vasculature and the Lung

Abstract

Electroporation has proven to be a highly effective technique for the in vivo delivery of genes to a number of solid tissues. In most of the reported methods, DNA is injected into the target tissue and electrodes are placed directly on or in the tissue for application of the electric field. While this works well for solid tissues, there are many tissues and organs that are not amenable to such an approach. In this review I will focus on the development of electroporation protocols for two such tissues: the vasculature and the lung. Several methods for in vivo electroporation of the vasculature have been developed in recent years that deliver DNA to vessel segments from either the inside or outside of the vessel. The advantages and disadvantages of each are discussed, as are the applications for which they have been used. In more recent work, our laboratory has developed a novel method to deliver genes to the rodent lung that results in high level, uniform, gene expression throughout all cell types of the lung. Most importantly, this technique is safe, and causes no inflammatory response or alterations in normal physiology of the organs. Taken together, these studies demonstrate the utility of electroporation for gene transfer to noninjectible tissues.

Nonviral gene transfer to skeletal, smooth, and cardiac muscle in living animals

“…Indeed, the physiological response was equivalent to that seen with recombinant adenoviruses expressing the same gene product (35,75). Most importantly, gene expression was detected throughout all cell types in the lung (21,23,75). Cell types that received and expressed DNA included alveolar type I and type II epithelial cells, endothelial cells, airway epithelial cells, and vascular and airway SMCs.…”

“…Airway smooth muscle also suffers from the fact that it is buried beneath other cells and is hidden from standard gene transfer reagents. Our lab has recently shown (21,23,75) that electroporation can also be used to target these cells without inducing any injury to the airways to make the cells accessible. Purified plasmid suspended in a physiologically compatible buffer containing 140 mM NaCl is administered to the airways of anesthetized mice or rats and the animals are electroporated with flat electrodes placed on either side of the chest.…”

“…By immunohistochemistry for the expressed gene product, there appeared to be little to no difference between the levels of gene expression in the various cell types. As for uniformity of gene transfer, there appears to be transfer and expression to all regions of the lung that is relatively homogeneous over a two-to threefold range (21,75).…”

Properties of WT and mutant hERG K+ channels expressed in neonatal mouse cardiomyocytes

“…cDNA (3-5 g; WT, ⌬Y475, G601S, or N470D hERG or empty pCDNA3 vector) and 1 g cDNA of green fluorescent protein (GFP) were transfected into neonatal mouse cardiomyocytes by an electroporation technique (7,13,30) using a Rat Cardiomyocyte-Neonatal Nucleofection Kit (Amaxa Biosystems, Cologne, Germany). HEK-293 cells were transfected using SuperFect (Qiagen) as previously described (3).…”

Transdermal Drug Delivery and Methods to Enhance It

“…Способ электропорации сделал возможным чрескожную доставку лекарств при воспалительных заболевани-ях [49]; обезболивании [50], химиотерапии опухо-лей [51][52][53][54][55][56][57]. Кроме того, электропорация расширяет возможности генной терапии, позволяющий введе-ние ДНК, генов в клетки [58,59], а также для вак-цин [60][61][62].…”