INTRODUCTION OF PLASMID DNA AND NUCLEAR-PROTEIN INTO CELLS BY USING ERYTHROCYTE-GHOSTS, LIPOSOMES, AND SENDAI VIRUS

This chapter focuses on developing the methods first to introduce exogenous DNA into the cytoplasm efficiently by Sendai virus (hemagglutinating virus of Japan, HVJ)-mediated fusion, and then to transfer the DNA efficiently from the cytoplasm to the nucleus for high level expression even in cells in the G O phase. for this purpose, new vehicles by using HVJ are developed to combine DNA-loaded liposomes with RBC ghosts containing nuclear proteins. by use of these vehicles, referred to here as vesicle complexes, DNA and nuclear proteins could be simultaneously introduced into cultured cells at more than 95% efficiency and the introduced DNA rapidly migrated into the nucleus even of interphase cells. by this delivery system, exogenous DNA was introduced into adult rat liver and highly but transiently expressed in hepatocytes in vivo. Liposomes containing DNA and proteins are incubated with HVJ, and the resulting HVJ-liposomes can introduce DNA and proteins simultaneously into the same cells as efficiently as the vesicle complex, and are also available for in vivo experiments. © 1993