Human rhinovirus internal ribosome entry site element enhances transgene expression in transfected CHO-S cells.
Human rhinovirus internal ribosome entry site element enhances transgene expression in transfected CHO-S cells.
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人鼻病毒内部核糖体进入位点元件增强转染 CHO-S 细胞中的转基因表达
DOI：
10.1038/s41598-018-25049-9
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发表时间：
2018-04-27
影响因子：
4.6
通讯作者：
Wang TY
中科院分区：
文献类型：
--
作者：
Chai YR;Ge MM;Wei TT;Jia YL;Guo X;Wang TY
Chinese hamster ovary (CHO) cells are mainly used for recombinant protein production. However, the unstable transgene expression and lower transgene copy numbers are the major issues need to be resolved. Here, eleven internal ribosome entry site (IRES) elements from viral and cellular IRES were evaluated for foreign gene expression in CHO-S cells. We constructed eleven fusing plasmids containing different IRES sequences downstream of the enhanced green fluorescent protein (EGFP) gene. EGFP expression was detected by flow cytometry and the transgene copy number was evaluated by quantitative PCR. The erythropoietin (EPO) protein was also used to assess the stronger IRES. The results showed that IRES from human rhinovirus (HRV) exhibited the highest EGFP expression level under transient and stable transfections. The EGFP expression level of vector with IRES from HRV was related to the gene copy number in stably transfected CHO-S cells. Moreover, IRES from HRV induced higher expression level of EPO compared with one mutant IRES from EMCV in transfected cells. In conclusion, IRES from HRV can function as a strong IRES element for stable expression in CHO-S cells, which could potentially guide more effective foreign gene expression in CHO-S cells.
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