Fusion with matrix attachment regions enhances expression of recombinant protein in human HT-1080 cells.

Autor: Jing CQ; Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Xinxiang 453003, Henan, China; International Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang 453003, Henan, China., Guo ML; Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Xinxiang 453003, Henan, China; International Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang 453003, Henan, China., Wang C; School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, China., Ni TJ; Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Xinxiang 453003, Henan, China; International Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang 453003, Henan, China., Guo X; Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Xinxiang 453003, Henan, China; International Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang 453003, Henan, China., Wang TY; Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Xinxiang 453003, Henan, China; International Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang 453003, Henan, China. Electronic address: wtianyuncn@126.com.
Jazyk: angličtina
Zdroj: Journal of bioscience and bioengineering [J Biosci Bioeng] 2020 Nov; Vol. 130 (5), pp. 533-538. Date of Electronic Publication: 2020 Aug 06.
DOI: 10.1016/j.jbiosc.2020.07.007
Abstrakt: Like endogenous proteins, recombinant foreign proteins produced in human cell lines also need post-translational modifications. However, high and long-term expression of a gene of interest (GOI) presents significant challenges for recombinant protein production in human cells. In this work, the effect of human matrix attachment region elements (MARs), including the β-globin MAR (gMAR), chicken lysozyme MAR (cMAR), and a combination of these two, on the stable expression of GOI was assessed in human HT-1080 cells. After transfection with vectors containing the MAR elements and eGFP, stably HT-1080 cell pools were obtained under selective pressure. eGFP protein expression was analyzed by flow cytometry, while transgene copy number and eGFP mRNA expression levels were determined with qPCR and qRT-PCR technology. We found that MARs could not enhance transfection efficiency, but gMAR could significantly increase eGFP expression in stable HT-1080 cell pools by approximately 2.69-fold. Moreover, gMAR could also increase eGFP expression stability during long-term culture. Lastly, we showed that the effect of the MARs on transgenes was related to the gene copy number. In summary, this study found that MARs could both enhance the transgene expression and stability in HT-1080 cells.
(Copyright © 2020 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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