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 language | English |
|---|---|
| Pages (from-to) | 621-630 |
| Number of pages | 10 |
| Journal | Molecular Biotechnology |
| Volume | 56 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - 2014 |
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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 journal › Article
}
TY - JOUR
T1 - Dramatic increase in expression of a transgene by insertion of promoters downstream of the cargo gene
AU - Sakaguchi, Masakiyo
AU - Watanabe, Masami
AU - Kinoshita, Rie
AU - Kaku, Haruki
AU - Ueki, Hideo
AU - Futami, Junichiro
AU - Murata, Hitoshi
AU - Inoue, Yusuke
AU - Li, Shun Ai
AU - Huang, Peng
AU - Putranto, Endy Widya
AU - Ruma, I. Made Winarsa
AU - Nasu, Yasutomo
AU - Kumon, Hiromi
AU - Huh, Nam Ho
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
KW - Adenovirus
KW - Gene expression
KW - Gene therapy
KW - Plasmid
KW - Recombinant protein
UR - http://www.scopus.com/inward/record.url?scp=84904347159&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84904347159&partnerID=8YFLogxK
U2 - 10.1007/s12033-014-9738-0
DO - 10.1007/s12033-014-9738-0
M3 - Article
C2 - 24526517
AN - SCOPUS:84904347159
VL - 56
SP - 621
EP - 630
JO - Molecular Biotechnology
JF - Molecular Biotechnology
SN - 1073-6085
IS - 7
ER -