In vivo gene transfer using cetylated polyethylenimine

Area of Science:

• Biotechnology
• Gene Therapy
• Nanomedicine

Background:

• Polyethylenimine (PEI) is a common non-viral gene delivery vector.
• Low-molecular-mass PEI (600 Da) requires modification for improved efficacy.
• Cetylation of PEI enhances its properties for gene delivery.

Purpose of the Study:

• To evaluate cetylated low-molecular-mass polyethylenimine (CT-PEI) formulated in liposomes for gene transfer.
• To assess the protective effect of CT-PEI liposomes against serum-induced DNA degradation.
• To compare the in vitro and in vivo gene expression efficiency of different CT-PEI liposome formulations.

Main Methods:

• Synthesis and characterization of CT-PEI (600 Da, 28% cetyl substitution).
• Formulation of CT-PEI into liposomes with cholesterol or dioleoylphosphatidylcholine.
• In vitro serum-induced plasmid DNA degradation assay.
• In vitro luciferase gene expression assay in serum-containing medium.
• In vivo systemic gene transfer in mice, assessing luciferase expression in lungs and liver.

Main Results:

• CT-PEI-containing liposomes protected complexed DNA from serum degradation.
• In vitro, CT-PEI/cholesterol liposomes yielded the highest luciferase gene expression.
• In vivo systemic gene transfer was most efficient with CT-PEI/cholesterol liposomes.
• Higher luciferase expression was observed in mouse lungs compared to the liver.

Conclusions:

• Liposomes containing cetylated polyethylenimine and cholesterol are a suitable vehicle for investigating systemic plasmid DNA transfer into lungs.
• CT-PEI/cholesterol liposomes offer a promising platform for targeted gene delivery to pulmonary tissues.
• This formulation demonstrates potential for therapeutic applications requiring efficient lung gene transfer.