A novel regulatory element (E77) isolated from CHO-K1 genomic DNA enhances stable gene expression in Chinese hamster ovary cells.
A novel regulatory element (E77) isolated from CHO-K1 genomic DNA enhances stable gene expression in Chinese hamster ovary cells.
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DOI：
10.1002/biot.201500464
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发表时间：
2016-05
影响因子：
4.7
通讯作者：
Lee, Eun Gyo
中科院分区：
文献类型：
--
作者：
Kang, Shin-Young;Kim, Yeon-Gu;Kang, Seunghee;Lee, Hong Weon;Lee, Eun Gyo
关键词：
Vectors flanked by regulatory DNA elements have been used to generate stable cell lines with high productivity and transgene stability; however, regulatory elements in Chinese hamster ovary (CHO) cells, which are the most widely used mammalian cells in biopharmaceutical production, are still poorly understood. We isolated a novel gene regulatory element from CHO‐K1 cells, designated E77, which was found to enhance the stable expression of a transgene. A genomic library was constructed by combining CHO‐K1 genomic DNA fragments with a CMV promoter‐driven GFP expression vector, and the E77 element was isolated by screening. The incorporation of the E77 regulatory element resulted in the generation of an increased number of clones with high expression, thereby enhancing the expression level of the transgene in the stable transfectant cell pool. Interestingly, the E77 element was found to consist of two distinct fragments derived from different locations in the CHO genome shotgun sequence. High and stable transgene expression was obtained in transfected CHO cells by combining these fragments. Additionally, the function of E77 was found to be dependent on its site of insertion and specific orientation in the vector construct. Our findings demonstrate that stable gene expression mediated by the CMV promoter in CHO cells may be improved by the isolated novel gene regulatory element E77 identified in the present study.
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影响因子：
2.2
作者：
Huang, Ying;Li, Yan;Shen, Bei Fen
通讯作者：
Shen, Bei Fen
影响因子：
14.9
作者：
Cho Y;Song SH;Lee JJ;Choi N;Kim CG;Dean A;Kim A
通讯作者：
Kim A
DOI：
10.1007/978-1-61779-352-3_9
发表时间：
2012-01-01
期刊：
PROTEIN EXPRESSION IN MAMMALIAN CELLS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Mariati;Ho, Steven C. L.;Yang, Yuansheng
通讯作者：
Yang, Yuansheng
影响因子：
3.8
作者：
Brown, Adam J.;Sweeney, Bernie;James, David C.
通讯作者：
James, David C.
影响因子：
2.9
作者：
Aldrich, TL;Viaje, A;Morris, AE
通讯作者：
Morris, AE