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Journal of Virology, July 2000, p. 6564-6569, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Cytoplasmic RNA Vector Derived from Nontransmissible Sendai Virus with Efficient Gene Transfer and Expression

Hai-Ou Li,1 Ya-Feng Zhu,1 Makoto Asakawa,1 Hidekazu Kuma,1 Takahiro Hirata,1 Yasuji Ueda,1 Yun-Sik Lee,1 Masayuki Fukumura,1 Akihiro Iida,1,* Atsushi Kato,2,dagger Yoshiyuki Nagai,2,Dagger and Mamoru Hasegawa1

DNAVEC Research Inc., Tsukuba-shi, Ibaraki 305-0856,1 and Department of Viral Infection, Institute of Medical Sciences, University of Tokyo, Minato-ku, Tokyo 108-8639,2 Japan

Received 18 January 2000/Accepted 6 April 2000

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 × 108 to 1.0 × 108 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.

* Corresponding author. Mailing address: DNAVEC Research Inc., 1-25-11 Kannondai, Tsukuba-shi, Ibaraki 305-0856, Japan. Phone: 81-298-38-0540. Fax: 81-298-39-1123. E-mail: iida{at}dnavec.co.jp.

dagger Present address: Department of Viral Diseases and Vaccine Control, National Institute of Infectious Diseases, Tokyo 208-0011, Japan.

Dagger Present address: AIDS Research Center, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.

Journal of Virology, July 2000, p. 6564-6569, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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