Dramatic increase in expression of a transgene by insertion of promoters downstream of the cargo gene.
Dramatic increase in expression of a transgene by insertion of promoters downstream of the cargo gene.
复制标题
DOI：
10.1007/s12033-014-9738-0
复制
发表时间：
2014-07
影响因子：
2.6
通讯作者：
Huh, Nam-ho
中科院分区：
文献类型：
--
作者：
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
为了在哺乳动物细胞中表达基因，已经开发了使用包括CMV、EF-1α和CAG启动子的启动子的各种载体，并且已经广泛使用。然而，根据货物基因的性质和/或宿主细胞类型，此类表达载体有时不能达到足够的表达水平。在本研究中，我们的目的是开发一种有效的启动子系统，使货物基因的高表达水平，在许多不同的细胞类型无处不在。我们发现，插入一个额外的启动子下游的货物基因大大提高了表达水平。在我们测试的构建体中，C-TSC盒（C：CMV-RU 5 '位于上游; TSC：由三重串联启动子、hTERT、SV 40和CMV组成的另一启动子单元，位于cDNA下游加上聚腺苷酸化信号）具有最有效的能力，显示出比有效的常规载体系统高得多的效率。结果表明，新的表达系统是有用的哺乳动物细胞中的重组蛋白的生产和应用作为一种基因治疗措施。
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.
登录
查看更多内容
影响因子：
3.7
作者：
Norrman K;Fischer Y;Bonnamy B;Wolfhagen Sand F;Ravassard P;Semb H
通讯作者：
Semb H
影响因子：
11.4
作者：
NEUHAUS, G;NEUHAUSURL, G;SCHWEIGER, HG
通讯作者：
SCHWEIGER, HG
影响因子：
64.5
作者：
BOSHART, M;WEBER, F;SCHAFFNER, W
通讯作者：
SCHAFFNER, W
影响因子：
6.4
作者：
Kawasaki, K.;Watanabe, M.;Date, H.
通讯作者：
Date, H.
影响因子：
3.5
作者：
NIWA, H;YAMAMURA, K;MIYAZAKI, J
通讯作者：
MIYAZAKI, J