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Nucleic Acids Research, 2002, Vol. 30, No. 8 1808-1816
© 2002 Oxford University Press

Synergy between cationic lipid and co-lipid determines the macroscopic structure and transfection activity of lipoplexes

Marilyn E. Ferrari, Denis Rusalov, Joel Enas and Carl J. Wheeler*

Department of Chemistry, Vical Incorporated, 9373 Towne Centre Drive, San Diego, CA 92121, USA

The large number of cytofectin and co-lipid combinations currently used for lipoplex-mediated gene delivery reflects the fact that the optimal cytofectin/co-lipid combination varies with the application. The effects of structural changes in both cytofectin and co-lipid were systematically examined to identify structure–activity relationships. Specifically, alkyl chain length, degree of unsaturation and the head group to which the alkyl side chain was attached were examined to determine their effect on lipoplex structure and biological activity. The macroscopic lipoplex structure was assessed using a dye-binding assay and the biological activity was examined using in vitro transfection in three diverse cell lines. Lipoplexes were formulated in three different vehicles currently in use for in vivo delivery of naked plasmid DNA (pDNA) and lipoplex formulations. The changes in dye accessibility were consistent with structural changes in the lipoplex, which correlated with alterations in the formulation. In contrast, transfection activity of different lipoplexes was cell type and vehicle dependent and did not correlate with dye accessibility. Overall, the results show a correlation between transfection and enhanced membrane fluidity in both the lipoplex and cellular membranes.

* To whom correspondence should be addressed. Tel: +1 858 646 1216; Fax: +1 858 646 1250; Email: cwheeler{at}vical.com


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