Cell-Specific Targeting Strategies for Electroporation-Mediated Gene Delivery in Cells and Animals
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Cell-Specific Targeting Strategies for Electroporation-Mediated Gene Delivery in Cells and Animals David A. Dean
Received: 19 December 2012 / Accepted: 8 March 2013 Ó Springer Science+Business Media New York 2013
Abstract The use of electroporation to facilitate gene transfer is an extremely powerful and useful method for both in vitro and in vivo applications. One of its great strengths is that it induces functional destabilization and permeabilization of cell membranes throughout a tissue leading to widespread gene transfer to multiple cells and cell types within the electric field. While this is a strength, it can also be a limitation in terms of cell-specific gene delivery. The ability to restrict gene delivery and expression to particular cell types is of paramount importance for many types of gene therapy, since ectopic expression of a transgene could lead to deleterious host inflammatory responses or dysregulation of normal cellular functions. At present, there are relatively few ways to obtain cell-specific targeting of nonviral vectors, molecular probes, small molecules, and imaging agents. We have developed a novel means of restricting gene delivery to desired cell types based on the ability to control the transport of plasmids into the nuclei of desired cell types. In this article, we discuss the mechanisms of this approach and several applications in living animals to demonstrate the benefits of the combination of electroporation and selective nuclear import of plasmids for cell-specific gene delivery. Keywords Electroporation Gene delivery Nuclear import Nucleus Trafficking Transfection
D. A. Dean (&) Departments of Pediatrics and Biomedical Engineering, University of Rochester, 601 Elmwood Avenue, Rochester, NY 14642, USA e-mail: [email protected]
Electroporation uses electrical fields to create transient pores in the cell membrane that allow the entry of normally impermeable macromolecules into the cytoplasm (Escoffre et al. 2009). While this technique is used most commonly to transfer DNA to bacteria, yeast, and mammalian cells in culture, it also can be applied very effectively to living animals. In most animal studies, electroporation causes anywhere from a 20- to a 1,000-fold increase in gene expression compared to DNA injection alone in normal tissues such as skin, liver, and muscle, as well as in a variety of tumors (Cemazar et al. 2009; Heller et al. 2000; Lin et al. 2012; Wells et al. 2000). We and others have adapted this technique for use in the vasculature (Martin et al. 2000; Young et al. 2003; Young et al. 2008), cornea (Blair-Parks et al. 2002), lung (Dean et al. 2003; MachadoAranda et al. 2005; Mutlu et al. 2007; Zhou et al. 2007), heart (Aistrup et al. 2009; Hargrave et al. 2013; Marshall et al. 2010), skin (Heller et al. 2002, 2010), skeletal muscle (Mir et al. 1998, 1999), liver (Heller et al. 1996), intestine, kidney, and prostate (Dean, unpublished), among other organs. The procedure is rapid, safe, and reproducible. Perhaps most exciting about this appro
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