Inducible vesicular stomatitis virus (VSV) L cell line for packaging of recombinant VSV.
Virus Genes
; 31(2): 195-201, 2005 Oct.
Article
en En
| MEDLINE
| ID: mdl-16025245
Recently, recombinant vesicular stomatitis viruses (VSV) have been developed as high-level expression vectors which serve as effective vaccine vectors in animals. An ideal approach for VSV vector production would be the development of stable packaging cell lines that can produce vector particles without transfection step. In this report, we describe generation of an inducible cell line that expresses the VSV polymerase gene (L) under the control of the reverse tetracycline-controlled transactivator (rtTA) system as a first step to make VSV-based packaging cell lines. Integrated polymerase (L) gene was controlled by an rtetR-dependent promoter in the rtTA-producing BHK cell line. When the cell lines were cultured in the presence of tet (tetracycline) or tetracycline derivative doxycycline, the recombinant VSV and wild type VSV were replicated, whereas in the absence of tet or tetracycline derivative doxycycline, the recombinant VSV was not replicated. Viral supernatants were harvested, diluted, and monitored by plaque assay for the presence of infectious VSV. Plaques of VSV containing an additional sequence encoding the EGFP protein allowed rapid detection of infection. Our results suggest wide applications of other surrogate viruses based on VSV. The availability of stable packaging cell lines represents a step toward the use of a VSV vector delivery system that can allow scale-up production of vector-stocks for gene therapy.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Proteínas Virales
/
ARN Polimerasa Dependiente del ARN
/
Ingeniería Genética
/
Virus de la Estomatitis Vesicular Indiana
/
Ensamble de Virus
/
Vectores Genéticos
Límite:
Animals
Idioma:
En
Revista:
Virus Genes
Asunto de la revista:
BIOLOGIA MOLECULAR
/
VIROLOGIA
Año:
2005
Tipo del documento:
Article
Pais de publicación:
Estados Unidos