Dramatic increase in expression of a transgene by insertion of promoters downstream of the cargo gene

Masakiyo Sakaguchi, Masami Watanabe, Rie Kinoshita, Haruki Kaku, Hideo Ueki, Junichiro Futami, Hitoshi Murata, Yusuke Inoue, Shun Ai Li, Peng Huang, Endy Widya Putranto, I. Made Winarsa Ruma, Yasutomo Nasu, Hiromi Kumon, Nam Ho Huh

Research output: Contribution to journalArticle

30 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)621-630
Number of pages10
JournalMolecular Biotechnology
Volume56
Issue number7
DOIs
Publication statusPublished - 2014

Fingerprint

Transgenes
Genes
Peptide Elongation Factor 1
Polyadenylation
Cells
Recombinant proteins
Recombinant Proteins
Complementary DNA
Gene Expression
Therapeutics

Keywords

  • Adenovirus
  • Gene expression
  • Gene therapy
  • Plasmid
  • Recombinant protein

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Molecular Biology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Medicine(all)

Cite this

Dramatic increase in expression of a transgene by insertion of promoters downstream of the cargo gene. / Sakaguchi, Masakiyo; Watanabe, Masami; Kinoshita, Rie; Kaku, Haruki; Ueki, Hideo; Futami, Junichiro; Murata, Hitoshi; Inoue, Yusuke; Li, Shun Ai; Huang, Peng; Putranto, Endy Widya; Ruma, I. Made Winarsa; Nasu, Yasutomo; Kumon, Hiromi; Huh, Nam Ho.

In: Molecular Biotechnology, Vol. 56, No. 7, 2014, p. 621-630.

Research output: Contribution to journalArticle

Sakaguchi, Masakiyo ; Watanabe, Masami ; Kinoshita, Rie ; Kaku, Haruki ; Ueki, Hideo ; Futami, Junichiro ; Murata, Hitoshi ; Inoue, Yusuke ; Li, Shun Ai ; Huang, Peng ; Putranto, Endy Widya ; Ruma, I. Made Winarsa ; Nasu, Yasutomo ; Kumon, Hiromi ; Huh, Nam Ho. / Dramatic increase in expression of a transgene by insertion of promoters downstream of the cargo gene. In: Molecular Biotechnology. 2014 ; Vol. 56, No. 7. pp. 621-630.
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