Cover Image

Not for Sale



View/Hide Left Panel

FIG. 1. Procedure of gene transfer by HVJ-liposome.

hr (15). In contrast, after direct transfer of fluorescein isothiocianate-ODN to VSMCs without the HVJ lipsome, fluorescence was observed in cytoplasmic compartments but not in the nucleus. Furthermore, the fluorescence was no longer detectable at 24 hr after direct fluorescein isothiocianateODN application.

Current gene transfer methods are also limited by the low level of expression of the transgene. We have found that cointroduction of plasmid DNA with a nuclear protein, high mobility group-1, can enhance transgene expression in animal tissues (16, 17). High mobility group-1 protein is a nonhistone DNA-binding protein of 28 kDa. It is reported that high mobility group-1 is required for the bending or looping of DNA and for enhancing transcription by specific recognition of cruciform DNA (18). An advantage of the HVJ-liposome is the capacity for such cointroduction of both DNA and proteins via their incorporation into the same particle. Indeed, we have recently cointroduced RNase H with antisense (AS) ODN for angiotensin converting enzyme in vivo into injured rat carotid artery. We observed that the AS effect was augmented 3-fold with the addition of RNase H (unpublished data). Thus, HVJ-liposomes have been useful for DNA transfer in various tissues in vivo, resulting in functional gene expression or gene suppression (Table 1).

Advantages of Fusigenic Viral Liposome. Efficient transfection of oligonucleotides, plasmid DNA, and proteins. The HVJ-liposome can encapsulate DNA up to 100 kbp. We have succeeded in transducing cosmid DNA (45 kbp) containing the thymidine kinase gene into cultured mouse cells (19). Recently, full-length cDNA of human Ducchene muscular dystrophy gene was introduced in vivo using HVJ-liposomes, resulting in its expression in skeletal muscle and diaphragm of the mdx mouse (20). When AS ODN to basic fibroblast growth factor was transfected into VSMCs using HVJ-liposomes and compared with cationic lipid transfection or direct ODN transfer without any vector, the concentration of AS basic fibroblast growth factor required to reduce cellular DNA synthesis by 75% was approximately 0.1 μM, 10 μM, and 20 μM, respectively (21). Thus, the HVJ-liposome is an effective method for ODN and plasmid DNA transfer. Ribozymes have also been efficiently introduced into cells using HVJ-liposomes, and the vector has also been useful for the introduction of recombinant proteins IgG and IgM (ref. 20 and unpublished data).

Penetration of the vector into tissues in vivo. Efficient in vivo transfer and expression of transgenes have been observed in cells of the tunica media of intact rabbit carotid arteries after filling the lumen with HVJ-liposomes containing the trans-

Table 1. In vivo gene transfer by HVJ-liposome

Organ

Gene product

Duration of gene expression

Liver

Rat and mouse

Insulin

7–14 days

Rat

Renin, HBsAg

7 days

Rat

LacZ

>4 weeks

Kidney

Rat

TGF-β, PDGF SV40-large tag

7 days

Heart

Rat

TGF-β, HSP70, Mn-SOD, Bcl-2

>2 weeks

>2 weeks

Skeletal muscle

Mouse

Dystrophin

2 weeks

Rat

Luciferase

>4 weeks

Rat

Decorin

>2 weeks

Artery

Rat

SV40, ACE, c-NOS, p21, ANP

>2 weeks

Rabbit

p53

>2 weeks

Lung

Rat

TGF-β, PDGF, LacZ

>2 weeks

Patellar ligament

Rat

LacZ

>4 weeks

Brain

Rat

LacZ

>2 weeks

Eye

Mouse and Monkey

LacZ

>2 weeks

Skin

Rat

LacZ

7–10 days

Testis

Mouse

CAT

>8 months

HBsAg, hepatitis B surface antigen; TGF-β, transforming growth factor β; PDGF, platelet-derived growth factor; SV40, simian virus 40; HSP70, heat shock proteins 70; SOD, superoxide dismutase; ACE, angiotensin converting enzyme; c-NOS, constitutive NO synthase; ANP, atrial natriuretic protein; and CAT, chloramphenicol acetyltransferase.



The National Academies | 500 Fifth St. N.W. | Washington, D.C. 20001
Copyright © National Academy of Sciences. All rights reserved.
Terms of Use and Privacy Statement