Nonviral Gene Transfer

Less invasive methods of delivering nonviral vectors to the CNS would be more desirable. Despite the minimal inflammatory profile of intracranial delivery of DNA-lipid complexes, the potential exists for surgery-induced adverse events, while the relatively limited volume of cUstribution obtained from stereo tac tic infusion makes the approach rather infeasible when attempting to treat cUseases that involve expansive regions of the brain. To that end, methodologies have been developed to enable the injection of mocUfied 

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Fabre, V., Boutrel, B., Hanoun, N. el al. (2000). Homeostatic regulation of serotonergic function by the serotonin transporter as revealed by nonviral gene transfer. J. Neurosci 20, 5065-75. 

BrunnerS, FurtbauerE, Sauer T, KursaM, Wagner E (2002) Overcoming the nuclear barrier cell cycle independent nonviral gene transfer with linear polyethylenimine or electroporation. Mol Ther 5 80-86... 

Cartier R, Reszka R. Utilization of synthetic peptides containing nuclear localization signals for nonviral gene transfer systems. J Biol Chem 2002 9(3) 157-167. 

The simplest nonviral gene transfer system in use for gene therapy is the injection of naked plasmid DNA (pDNA) into local tissues or the systemic circulation (88, 100). Naked DNA systems are composed of a bacterial plasmid that contains the cDNA of a reporter or therapeutic gene under the transcriptional control of various regulatory elements (101, 102). In recent years, work in several laboratories has shown that naked plasmid DNA (pDNA) can be delivered efficiently to cells in vivo either via electroporation, or by intravascular delivery, and has great prospects for basic research and gene therapy (101). Efficient transfection levels have also been obtained on direct application of naked DNA to the liver (103, 104), solid tumours (105), the epidermis (106), and hair follicles (106). 

W. C. Heiser, Ed., Gene Delivery to Mammalian Cells. Vol. 1. Nonviral Gene Transfer Techniques, Totowa, NJ Humana, 2004. 

Pitkanen L, Ruponen M, Nieminen J, Urtti A. Vitreous is a barrier in nonviral gene transfer by cationic lipids and polymers. Pharm Res 2003 20(4) 576-583. 

They also investigated the localization of arginine peptide/DNA complexes in the nucleus. Earlier studies showed that there was a major barrier in nuclear transportation for nonviral gene transfer [107,108]. They used double labeled complexes with rhodamine and FITC for the determination of the nuclear transport of arginine peptide/DNA complexes. They observed the fluorescence predominantly on the extracellular surface after Ihr incubation and after 2 hr incubation, the fluorescence was found in the cytoplasm, which indicated the entry of the complexes into the cells. After 6 hr incubation, they observed the fluorescence in a perinuclear location and finally, the fluorescence was observed within the nucleus after 12 hr incubation. From this observation, they concluded that the arginine peptide/DNA complexes were transferred into the cell nucleus without the dissociation of the arginine peptide. 

Nonviral Gene Transfer Systems in Somatic Gene Therapy... 

Fig. 14.20. a-CyD/polyamidoamine dendrimer conjugate as a nonviral gene transfer vector. 

In 1970, Nenitzescu passed away, and Balaban continued working on pyrylium salts both at the Bucharest Polytechnic where he had become a professor, and at the Institute of Atomic Physics Bucharest, where he was head of the Laboratory of Isotopically Labeled Organic Compounds. By having a large collection of pyrylium salts, he reported on new reactions and physical properties of pyrylium salts. More recently, he prepared from pyrylium salts new pyridinium salts that were designed to be ionic liquids or nonviral gene transfer agents. 

Small positively charged particles are formed by the self assembly of cationic, hydrolytically degradable poly(-amino esters) and plasmid DNA. These nanoparticles are biodegradable and suitable for nonviral gene transfer to stem cells moreover the end group of the polymer can be modified leading to improvement of the gene delivery efficacy [195]. 

Nonviral gene transfer systems are based on a variety of technologies that employ physical/chemical means to deliver genes [6], These technologies include direct plasmid injection, bombardment with DNA coated microprojectiles, and DNA complexed with liposomes or polymers. Some nonviral transfection techniques are too inefficient (e.g., coprecipitation of DNA with calcium phosphate [7], DNA complexed with diethylaminoethyl (DEAE)-dextran [8], electroporation [9]), or laborious (e.g., microinjection of DNA [ 10 ]) for clinical use. Only those gene delivery systems (viral and nonviral) with potential for clinical application are discussed in this chapter. The main features of these technologies (Table 18.3) are described and specific examples of their applications highlighted. 

With respect to nonviral gene transfer, strategies aimed at improving gene transfer efficiency include the ongoing modification of lipid vectors currently in use. Third-generation synthetic vectors have been designed, and they include ... 

We examined the characteristics of FL as in comparison with CL-DNA complex, a common nonviral gene transfer vector. 

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