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Oligonucleotide-cationic liposome interactions. A physicochemical study

I Jääskeläinen¹, J Mönkkönen, A Urtti

¹Department of Pharmaceutical Technology, University of Kuopio, Finland.

Biochimica Et Biophysica Acta

|October 12, 1994

Summary

Cationic liposomes and antisense oligonucleotides form complexes that aggregate and fuse. Fusion requires phosphatidylethanolamine or negative charge in cell membranes, with lipid ratio being critical.

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Area of Science:

Biochemistry
Materials Science
Drug Delivery

Background:

Cationic liposomes effectively deliver antisense oligonucleotides into cells.
Understanding the interactions between cationic lipids and oligonucleotides is crucial for optimizing delivery systems.

Purpose of the Study:

To investigate the aggregation and fusion reactions of cationic lipid/oligonucleotide complexes.
To determine the influence of lipid composition and charge ratio on complex formation and membrane interactions.
To elucidate the requirements for fusion with lipid bilayers.

Main Methods:

Complexation of phosphorothioate oligonucleotides with cationic liposomes (DDAB/DOPE and DOTAP).
Size analysis of complexes at varying charge ratios.
Resonance energy transfer experiments to study aggregation and fusion kinetics.

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Leakage assays using fluorescent dyes (calcein) with different liposome compositions.

Main Results:

Oligonucleotide-induced aggregation significantly increased complex size at charge ratios of 0.4-2.0.
Oligonucleotides induced liposome fusion, with kinetics dependent on the initial aggregation step.
Fusion and aggregation were inhibited at high charge ratios due to electrostatic repulsion.
Liposome leakage was observed with DOTAP/oligonucleotide complexes under specific charge ratios and lipid compositions.

Conclusions:

The ratio of oligonucleotide to cationic lipid critically influences the physicochemical properties of the complexes.
Phosphatidylethanolamine or a negative charge on the cell membrane is necessary for the fusion of cationic liposome-oligonucleotide complexes.
These findings provide insights into the mechanism of cationic liposome-mediated gene delivery.