The improved efficient method for introducing macromolecules into cells using HVJ (Sendai virus) liposomes with...

Area of Science:

• Molecular Biology
• Cell Biology
• Biotechnology

Background:

• Liposomes are widely used for drug and gene delivery.
• Efficient delivery of macromolecules into cells remains a challenge.
• Gangliosides can enhance liposome-mediated cellular uptake.

Purpose of the Study:

• To evaluate the efficiency of ganglioside-associated liposomes combined with HVJ (Sendai virus) for macromolecule delivery into cultured cells.
• To demonstrate gene transfer and stable transformation using this method.
• To assess the potential for therapeutic applications.

Main Methods:

• Macromolecules (DNA, RNA, genes) were entrapped in liposomes using reverse-phase evaporation.
• Liposomes were associated with gangliosides.
• Liposome-cell complexes were incubated with HVJ (Sendai virus).
• Gene transfer efficiency was assessed by measuring enzyme activity and obtaining transformants.
• Protein expression was analyzed via immunoprecipitation.

Main Results:

• HVJ liposomes with gangliosides achieved efficient delivery of DNA and RNA into cultured cells.
• High transient expression of thymidine kinase was observed in Ltk- cells (>95%).
• Stable transformants were obtained in various cell lines (L, CHO-Kl, HeLa-S3, Ehrlich ascites tumor cells) with frequencies up to 1.0%.
• Introduction of rat liver mRNA resulted in secretion of rat albumin and other proteins.
• Immunoglobulin M (IgM) was successfully introduced into all L cells.

Conclusions:

• HVJ liposomes incorporating gangliosides represent a highly efficient system for delivering macromolecules into cultured cells.
• This technology facilitates both transient gene expression and stable cellular transformation.
• The method shows significant potential for gene therapy and protein production applications.