Cationic lipid-mediated gene transfer: analysis of cellular uptake and nuclear import of plasmid DNA.

Abstract:

:Cationic lipids are widely used for gene transfer in vitro and show promise as vectors for in vivo gene therapy applications. However, there is limited understanding of the cellular mechanisms involved in nonviral gene transfer. We investigated two major steps that could be limiting barriers to cationic lipid-mediated gene transfer in vitro. We used a fluorescent plasmid to study the cellular uptake and the intracellular fate of lipoplexes during in vitro transfection of fibroblast cells and found that 100% of the cells take up lipoplexes. The intracellular staining observed with lipoplexes was clearly different from that obtained with endocytosed fluorescent dextran. This suggests that cells readily take up lipoplexes by a mechanism that could be different from endocytosis in our conditions. However, the escape of DNA from intracellular vesicles could be a major limiting barrier to gene transfer. Direct injection of plasmid DNA into the nucleus and cytoplasm of cells indicated that DNA traffic from the cytoplasm to the nucleus might be also an important limiting step.

journal_name

Cell Biol Toxicol

authors

Escriou V,Ciolina C,Helbling-Leclerc A,Wils P,Scherman D

doi

10.1023/a:1007425803756

subject

Has Abstract

pub_date

1998-03-01 00:00:00

pages

95-104

issue

2

eissn

0742-2091

issn

1573-6822

journal_volume

14

pub_type

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