Effect of co-lipids in enhancing cationic lipid-mediated gene transfer in vitro and in vivo

Abstract

Complexes of DNA and cationic lipids are promising vecuptake, whereas others had a greater effect on steps subtors for gene transfer. Most cationic lipid formulations consequent to entry. Based on those results, we tested the tain both a cationic component and a neutral co-lipid. We hypothesis that co-lipids conferring different properties found that the co-lipid could influence DNA uptake in COScould be combined to enhance gene transfer. The results 1 cells, but processes subsequent to uptake were even showed that a combination of co-lipids had a synergistic more important in determining gene expression. We effect on expression. We also found that structural analogs compared dioleoylphosphatidylethanolamine (DOPE) and of DOPE were more effective than DOPE in enhancing structural analogs of DOPE combined with cationic lipids gene transfer to mature human airway epithelia studied in and found that DNA uptake and transgene expression did vitro and to mouse lung studied in vivo. These data provide not always correlate. Transgene expression was depeninsight into the mechanism by which co-lipids influence catdent on DNA uptake into the cell, on entry of DNA into the ionic lipid-mediated gene transfer and show that optimizcytoplasm, and on release of DNA from the lipid complex.ation of the effects of co-lipids can enhance gene transfer We found that some co-lipids had a greater effect on DNA both in vitro and in vivo.

Transgene expression, but not gene delivery, is improved by adhesion-assisted lipofection of hematopoietic cells

“…The fact that DNA uptake and transgene expression do not always correlate in transfected cells, but that processes subsequent to DNA uptake could be even more important in determining gene expression, has been widely documented. 16,23 Thus, delivery of plasmid DNA to cultured epithelial cells was demonstrated not to be a limiting factor, since virtually 100% of cells receiving a lipid-DNA conjugate rapidly transport exogenous DNA to the region of the cell nucleus but only a small portion of these cells, are able to express the DNA at detectable levels. 24,25 We confirmed here that delivery of plasmid DNA into cultured cells would not be the only limiting factor, but would implicate features inherent in the cells as contributing to the poor efficiency of lipid-based gene expression observed in vitro.…”

“…8 The sequence of cellular and molecular events underlying the complex phenomenon of DNA transfer remains highly speculative, 9 but it is commonly accepted that endocytosis is the main mechanism of entry of liposome/DNA particles, [10][11][12][13] and relatively high transfection efficiency is due to the intrinsic membrane-rupturing capability of cationic liposomes as a result of destabilizing the plasma and/or endosome membrane. [14][15][16] In contrast to adherent cells, liposome/DNA particles have been reported to bind inefficiently to the cell surface of lymphocytes, suggesting that particles attach to membrane components involved in cell anchoring to the extracellular matrix and composed of cadherins or proteoglycans present on adherent cells only. 6 This weak interaction between particles and membrane lymphocytes would prevent endocytosis of liposome/DNA particles and release of DNA into the cytosol of nonadherent cells.…”

Folate-targeted, cationic liposome-mediated gene transfer into disseminated peritoneal tumors

“…In regards to gene delivery, the most common liposomes are those prepared with various cationic lipids in combination with a fusogenic lipid, such as DOPE. 35,36 For our investigation, we prepared FA-targeted liposomes where the folate moiety was attached to a lipid membrane anchor via a cysteinyl-PEG 3400 spacer. The Cys-PEG spacer was employed for two reasons.…”

Liposome-mediated NGF gene transfection following neuronal injury: potential therapeutic applications

“…It may be related to differences in cell surface composition that affect complex binding and differences in the mechanism of complex uptake by different cell types. 40 We have also found the incubation time can affect transgene product levels. Twelve hours of liposome-DNA complex incubation in cultures resulted in higher levels of NGF production than 6 h of incubation.…”