Human rhinovirus internal ribosome entry site element enhances transgene expression in transfected CHO-S cells

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 eval...

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Bibliographic Details
Published in:Scientific reports 2018-04, Vol.8 (1), p.6661-8, Article 6661
Main Authors: Chai, Yu-rong, Ge, Meng-meng, Wei, Ting-ting, Jia, Yan-long, Guo, Xiao, Wang, Tian-yun
Format: Article
Language:English
Subjects:
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Copy number
Erythropoietin
Flow cytometry
Gene expression
Green fluorescent protein
Humanities and Social Sciences
Internal ribosome entry site
multidisciplinary
Plasmids
Proteins
Rhinovirus
Science
Science (multidisciplinary)
Transgenes
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Summary: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.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-018-25049-9