title: Dramatic increase in expression of a transgene by insertion of promoters downstream of the cargo gene.
abstract: For expression of genes in mammalian cells, various vectors have been developed using promoters including CMV, EF-1α, and CAG promoters and have been widely used. However, such expression vectors sometimes fail to attain sufficient expression levels depending on the nature of cargo genes and/or on host cell types. In the present study, we aimed to develop a potent promoter system that enables high expression levels of cargo genes ubiquitously in many different cell types. We found that insertion of an additional promoter downstream of a cargo gene greatly enhanced the expression levels. Among the constructs we tested, C-TSC cassette (C: CMV-RU5' located upstream; TSC: another promoter unit composed of triple tandem promoters, hTERT, SV40, and CMV, located downstream of the cDNA plus a polyadenylation signal) had the most potent capability, showing far higher efficiency than that of potent conventional vector systems. The results indicate that the new expression system is useful for production of recombinant proteins in mammalian cells and for application as a gene therapeutic measure.
authors: Department of Cell Biology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, 700-8558, Japan, masa-s@md.okayama-u.ac.jp.
authors: Masakiyo
authors: M
authors: Sakaguchi
authors: Masami
authors: M
authors: Watanabe
authors: Rie
authors: R
authors: Kinoshita
authors: Haruki
authors: H
authors: Kaku
authors: Hideo
authors: H
authors: Ueki
authors: Junichiro
authors: J
authors: Futami
authors: Hitoshi
authors: H
authors: Murata
authors: Yusuke
authors: Y
authors: Inoue
authors: Shun-Ai
authors: SA
authors: Li
authors: Peng
authors: P
authors: Huang
authors: Endy Widya
authors: EW
authors: Putranto
authors: I Made Winarsa
authors: IM
authors: Ruma
authors: Yasutomo
authors: Y
authors: Nasu
authors: Hiromi
authors: H
authors: Kumon
authors: Nam-Ho
authors: NH
authors: Huh
mesh_terms: D000818:Animals; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D011401:Promoter Regions, Genetic; D019098:Telomerase / Q000096:biosynthesis* / Q000235:genetics; D019076:Transgenes
chemical_list: C509186:TERT protein, human; D019098:Telomerase