title: Development of high-throughput functional screening of therapeutic genes, using a hemagglutinating virus of Japan envelope vector.
abstract: Isolation of effective therapeutic genes is critical for the advancement of gene therapy for various diseases, including vascular diseases and cancers. The goal of the present study was to screen a human cDNA library, using a hemagglutinating virus of Japan envelope (HVJ-E) vector, to isolate candidate genes with potent therapeutic potential. The advantages of a high-throughput functional screening system based on the HVJ-E vector include (1) rapid preparation of the vector containing the DNA library, (2) effective fusion-mediated transfer of the plasmids to various cells with minimal toxicity, and (3) easy cloning of candidate genes by transformation of Escherichia coli. These advantages resulted in a lower probability of damage to isolated clones and in minimization of the time needed to screen for candidate genes. Screening of a human heart library for candidate genes to regulate endothelial cell growth identified three growth-stimulating genes, as evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and c-fos promoter activity, the products of which were more potent than vascular endothelial growth factor. Similarly, two growth-inhibiting genes were identified, the effects of which were similar to angiostatin. Overall, this novel system will help advance our understanding of cell biology and promote the utility of human gene therapy.
authors.affiliation: Division of Gene Therapy Science, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Nishikawa
authors.forename: Hironori
authors.initials: H
authors.lastname: Nakagami
authors.forename: Atsushi
authors.initials: A
authors.lastname: Matsuki
authors.forename: Akito
authors.initials: A
authors.lastname: Maeda
authors.forename: Chiah Yian
authors.initials: CY
authors.lastname: Yo
authors.forename: Taku
authors.initials: T
authors.lastname: Harada
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Morishita
authors.forename: Katsuto
authors.initials: K
authors.lastname: Tamai
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Kaneda
mesh_terms: D002460:Cell Line; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D004730:Endothelium, Vascular / Q000502:physiology; D015723:Gene Library; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000502:physiology; D029082:Sendai virus
chemical_list: D018076:DNA, Complementary