Confocal and probe microscopy to study gene transfection mediated by cationic liposomes with a cationic cholesterol derivative

A novel cationic cholesterol derivative with a hydroxyethyl amino head group (I) has been synthesized and used for liposome-mediated gene transfection. Cationic liposomes with derivative (I) greatly facilitated gene transfection into various cultured cells. The efficiency of transfection by liposomes with derivative (I) was much higher than that using liposomes with DC-chol (II) or lipofectine. Atomic force microscopy and confocal laser scanning microscopy revealed the molecular mechanism of gene transfection by cationic liposomes. The results showed that at least two steps were involved in gene transfection mediated by cationic liposomes. One was endocytosis, where the liposome–DNA complex was internalized into target cells, and the other was membrane fusion between the liposome vectors and endosomes, where DNA transferred from the liposome to the nucleus. In addition we found that microtubules were involved in the intracellular dynamics of gene transfection.