Influence of plasmid DNA topology on the transfection properties of DOTAP/DOPE lipoplexes

被引：88

作者：

Remaut, Katrien ^{[1
]}

Sanders, Niek N. ^{[1
]}

Fayazpour, Farzaneh ^{[1
]}

Demeester, Jozef ^{[1
]}

De Smedt, Stefaan C. ^{[1
]}

机构：

[1] Univ Ghent, Lab Gen Biochem & Phys Pharm, B-9000 Ghent, Belgium

来源：

JOURNAL OF CONTROLLED RELEASE | 2006年 / 115卷 / 03期

关键词：

plasmid DNA topology; DNA degradation; nuclear membrane; gene expression; DNA transfer; Transfection;

D O I：

10.1016/j.jconrel.2006.08.009

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Plasmid DNA (pDNA) can occur in the compact supercoiled (SC) form, the relaxed open circular (OC) form and the linearized form. In this paper we investigated the transfection efficiency of SC, OC and linearized pDNA complexed to DOTAP/DOPE liposomes in Vero cells. Only DOTAP/DOPE liposomes containing SC pDNA showed protein expression while DOTAP/DOPE liposomes loaded with OC or linearized pDNA failed. First we questioned if the better transfection properties of the SC pDNA-containing lipoplexes could be due to a better transcription of SC pDNA in the nuclei of the cells, compared to OC and linearized pDNA. However, microinjecting (naked) SC, OC or linearized pDNA in the nuclei of the Vero cells revealed that the transcription efficiency was independent on the pDNA topology but did depend on the intramiclear concentration of the pDNA. As the amount of pDNA that reaches the nucleus is determined by the amount of pDNA that arrives in the cytosol it could be hypothesized that SC pDNA is more efficiently released from the DOTAP/DOPE liposomes when compared to OC and linearized pDNA. However, microinjecting comparable concentrations of the pDNA topologies in the cytoplasm still resulted in a significantly higher transfection in the case of SC pDNA, especially in cells that underwent cell division in the period after injection. It seems that, compared to OC and linearized pDNA, SC pDNA is better suited to reach the perinuclear region, a prerequisite to become entrapped in the nuclei of the cells during cell division. (c) 2006 Elsevier B.V. All rights reserved.

引用

页码：335 / 343

页数：9

共 30 条

[1] Nuclear localization signal peptides enhance cationic liposome-mediated gene therapy [J].

Aronsohn, AI ;

Hughes, JA .

JOURNAL OF DRUG TARGETING, 1998, 5 (03) :163-169

[2]

Boulikas T, 1997, INT J ONCOL, V10, P301

[3] A peptide nucleic acid-nuclear localization signal fusion that mediates nuclear transport of DNA [J].

Brandén, LJ ;

Mohamed, AJ ;

Smith, CIE .

NATURE BIOTECHNOLOGY, 1999, 17 (08) :784-787

[4] Cell cycle dependence of gene transfer by lipoplex polyplex and recombinant adenovirus [J].

Brunner, S ;

Sauer, T ;

Carotta, S ;

Cotten, M ;

Saltik, M ;

Wagner, E .

GENE THERAPY, 2000, 7 (05) :401-407

[5] Effect of DNA topology on the transfection efficiency of poly((2-dimethylamino)ethyl methacrylate)-plasmid complexes [J].

Cherng, JY ;

Schuurmans-Nieuwenbroek, NME ;

Jiskoot, W ;

Talsma, H ;

Zuidam, NJ ;

Hennink, WE ;

Crommelin, DJA .

JOURNAL OF CONTROLLED RELEASE, 1999, 60 (2-3) :343-353

[6] Minicircle:: an improved DNA molecule for in vitro and in vivo gene transfer [J].

Darquet, AM ;

Rangara, R ;

Kreiss, P ;

Schwartz, B ;

Naimi, S ;

Delaère, P ;

Crouzet, J ;

Scherman, D .

GENE THERAPY, 1999, 6 (02) :209-218

[7] Actin cytoskeleton as the principal determinant of size-dependent DNA mobility in cytoplasm [J].

Dauty, E ;

Verkman, AS .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (09) :7823-7828

[8] Import of plasmid DNA into the nucleus is sequence specific [J].

Dean, DA .

EXPERIMENTAL CELL RESEARCH, 1997, 230 (02) :293-302

[9] Critical assessment of the nuclear import of plasmid during cationic lipid-mediated gene transfer [J].

Escriou, V ;

Carrière, M ;

Bussone, F ;

Wils, P ;

Scherman, D .

JOURNAL OF GENE MEDICINE, 2001, 3 (02) :179-187

[10] DNA transfection of macaque and murine respiratory tissue is greatly enhanced by use of a nuclease inhibitor [J].

Glasspool-Malone, J ;

Steenland, PR ;

McDonald, RJ ;

Sanchez, RA ;

Watts, TL ;

Zabner, J ;

Malone, RW .

JOURNAL OF GENE MEDICINE, 2002, 4 (03) :323-332

← 1 2 3 →