Polyethylenimine (PEI) is a simple, inexpensive and effective reagent for condensing and linking plasmid DNA to adenovirus for gene delivery

Abstract

A simple and inexpensive method of condensing and linktransfection complexes deliver plasmid DNA to cells by the ing plasmid DNA to carrier adenovirus particles is adenovirus infectious route without interference from virus described. The synthetic polycation polyethylenimine is gene expression because psoralen-inactivated virus is used to condense plasmid DNA into positively charged 100 employed. The PEI-DNA-adenovirus complexes display nm complexes. These PEI-DNA complexes are then DNA delivery comparable to more sophisticated DNA virus bound to adenovirus particles through charge interactions complexes employing streptavidin/biotin linkage, but require with negative domains on the viral hexon. The resulting no special reagents and are much easier to prepare.

Enhancement of calcium phosphate-mediated transfection by inclusion of adenovirus in coprecipitates

“…Figure 9 shows an evaluation of toxicity using the sensitive calcein AM, ethidium homodimer method and FACS analysis. 24,25 Following transfection of the cells for 1 h, the DNA:CaPi:Ad coprecipitates showed no increase in cell 14 and Lipofectin 8 were formed under optimal conditions based on previous reports and our own unpublished work and previous reports 1,13,14,23 …”

“…13,14 We chose these agents because they are widely used, are effective for transfection, and are available commercially. We generated complexes of plasmid DNA with each reagent and transfected NIH-3T3 cells.…”

“…A number of methods have been developed to facilitate plasmid-mediated gene transfer, including direct injection, electroporation, and incorporation of plasmid in complexes with cationic lipids, polylysine, and other molecules. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] The use of calcium phosphate (CaPi) to coprecipitate DNA was one of the first methods used for DNA transfer to cells. 16 CaPi, as well as cationic lipids and other cationic polymers, condense the DNA and facilitate its uptake into the cell.…”

“…A similar strategy has been described in which adenovirus was included in complexes of DNA plus a cationic molecule (polylysine or cationic lipids), and in some cases a targeting moiety. [5][6][7][8]10,12,14 In such strategies, the adenovirus was either covalently linked to the cationic molecule or included in the complex via electrostatic interactions. Including adenovirus in the complex by either method markedly improved delivery and expression of plasmid DNA.…”

Efficient adventitial gene delivery to rabbit carotid artery with cationic polymer–plasmid complexes

“…1,7,8 Cationic polymers have been widely used in gene transfer experiments in vitro and in vivo. [15][16][17][18][19][20][21] The purpose of the present study was to investigate the gene transfer efficiency of different liposomes and cationic polymers in vitro to smooth muscle cells (SMC) and endothelial cells (ECV 304) and to evaluate the most promising vehicles for in vivo gene delivery to rabbit carotid artery. Based on in vitro studies, fractured polyamidoamine dendrimers (generation 6) and PEI of 25 kDa and 800 kDa were chosen for in vivo study.…”