A cytoplasmic RNA vector derived from nontransmissible Sendai virus with efficient gene transfer and expression.
We have recovered a virion from defective cDNA of Sendai virus (SeV) that is capable of self-replication but incapable of transmissible-virion production. This virion delivers and expresses foreign genes in infected cells, and this is the first report of a gene expression vector derived from a defective viral genome of the Paramyxoviridae. First, functional ribonucleoprotein complexes (RNPs) were recovered from SeV cloned cDNA defective in the F (envelope fusion protein) gene, in the presence of plasmids expressing nucleocapsid protein and viral RNA polymerase. Then the RNPs were transfected to the cells inducibly expressing F protein. Virion-like particles thus obtained had a titer of 0.5 x 10(8) to 1. 0 x 10(8) cell infectious units/ml and contained F-defective RNA genome. This defective vector amplified specifically in an F-expressing packaging cell line in a trypsin-dependent manner but did not spread to F-nonexpressing cells. This vector infected and expressed an enhanced green fluorescent protein reporter gene in various types of animal and human cells, including nondividing cells, with high efficiency. These results suggest that this vector has great potential for use in human gene therapy and vaccine delivery systems.
Li
H O
HO
DNAVEC Research Inc., Tsukuba-shi, Ibaraki 305-0856, Japan.
Zhu
Y F
YF
Asakawa
M
M
Kuma
H
H
Hirata
T
T
Ueda
Y
Y
Lee
Y S
YS
Fukumura
M
M
Iida
A
A
Kato
A
A
Nagai
Y
Y
Hasegawa
M
M
D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D004730:Endothelium, Vascular / Q000166:cytology; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D007276:Injections, Intraventricular; D008099:Liver / Q000166:cytology; D008164:Luminescent Proteins / Q000235:genetics; D008168:Lung / Q000098:blood supply; D008253:Macaca mulatta; D009130:Muscle, Smooth / Q000166:cytology; D012367:RNA, Viral / Q000235:genetics*; D051381:Rats; D018108:Respirovirus / Q000235:genetics*
D008164:Luminescent Proteins; D012367:RNA, Viral; D049452:Green Fluorescent Proteins