Gene transfection by quantitatively reconstituted Sendai envelope proteins into liposomes

Abstract

Fusogenic liposomes ( virosomes ) consisting of Sendai virus envelope proteins have been utilized for in vitro and in vivo genetic modification of animal cells. In this study, the virosomes containing DNA were prepared by quantitative reconstitution of Sendai envelope proteins, fusion protein and hemagglutinin -neuramindase in liposomal vesicles. The Sendai virosomes more efficiently transferred genes into cultured 293 transformed kidney cells than 1,2 -dioleoyl -3 -( trimethylammonium ) propane -based cationic liposomes. At 200:1 weight ratio of envelope protein and lipid, the virosomes exhibited the best efficiency of gene transfection into the cells. The Sendai virosomes required relatively a short period of incubation time and much less cytotoxic, compared to the cationic liposome / DNA complex. The transfection efficiency of the Sendai virosomes containing DNA was maintained 70% after a month. This type of Sendai virosomes is relatively convenient for preparation and storage, compared to fusogenic liposomes prepared by liposome -virus fusion. First of all, because the constituents are quantitatively formulated, this type of virosome formulation can provide further consistent transfection for gene therapy.

Effect of Lipid Compositions on Gene Transfer into 293 Cells Using Sendai F/HN-virosomes

“…Recently we also reported that negatively-charged PS that contained Sendai virosomes efficiently transfer genes into cultured cells (Kim and Park, 2002). However, among the electron negatively-charged virosomes, the ASVE-and PSvirosomes were relatively efficient in gene transfer while the PA-virosomes were not (Fig.…”

“…The fusogenic proteins that were reconstituted into the virosomes were another major factor that governed transfection efficiency. In this study, the F/HN-proteins were reconstituted at a 1 : 100 weight ratio of protein to lipid, the ratio at which in vitro gene transfection has been found to be the most efficient (Kim and Park, 2002).…”

“…Tel: 82-33-760-2448;Fax: 82-33-763-5224 E-mail: yspark@dragon.yonsei.ac.kr Recently, we reported that the Sendai virosomes were an efficient vector for transferring the gene into 293 cells (Kim and Park, 2002). Virosomal constituents, such as the lipid components and their molar ratios, and the concentrations of viral envelope proteins can dramatically affect the efficiency of the gene transfer that is mediated by Sendai virosomes (Kaneda et al, 1999;Ponimaskin et al, 2000).…”

A Needle-Free Approach for Topical Immunization: Antigen Delivery via Vesicular Carrier System(s)

“…Moreover lipid vesicles also do not release the incorporated molecule instantaneously so they can act as local depot for sustained release of drug or immunogen. Some commonly employed vesicular carriers that facilitate permeation includes liposomes [5,63], niosomes [5,63,64], transfersomes [63,65], virosomes [66] Reconstituted Sandai Virus Envelop (RSVE) [69][70] etc. The driven forces, which are involved in selective transfer of lipid vesicles through intact skin, are osmotic gradient and hydration force generally.…”

An efficient liposomal gene delivery vehicle using Sendai F/HN proteins and protamine

“…15 We have developed a novel Sendai F/HN virosome system for in vivo gene transfection. The Sendai virosome formulations were optimized for in vitro and in vivo transfection, especially systemic gene delivery.…”