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Human genome-derived TOP1 matrix attachment region enhances transgene expression in the transfected CHO cells

  • Yan-Long Jia
  • Xiao Guo
  • Xi-Cheng Wang
  • Tian-Yun WangEmail author
Original Research Paper
  • 47 Downloads

Abstract

Objectives

To investigate the effect of full-length fragment of DNA topoisomerase I gene (TOP1) matrix attachment regions (MARs) originating from the human genome on transgene expression in Chinese hamster ovary (CHO) cells and explore the underlying mechanisms.

Results

Results showed that TOP1 MAR cannot only enhance the transient and stable transgenic expression of enhanced green fluorescence protein (EGFP) but also increase long-term stability and ratio of positive colonies in transfected CHO cells with TOP1 MAR at the 5′ or 3′ ends of the EGFP expression cassette. Interestingly, the CHO cells were transfected with the 5′,3′ TOP1 MAR-containing vector featured the highest transient and stable expression, whereas those with the 3′ TOP1 MAR-containing vector exhibited the most effective stability and ratio of positive colonies. We also observed that transgene copy numbers and mRNA of egfp gene were correlated with the expression levels of EGFP protein in polyclonal CHO cells. However, the heterogeneity of expression in monoclonal CHO cells was unaffected by transgene copy number.

Conclusions

The findings may aid in the potential application of TOP1 MAR in expression enhancement of recombinant proteins in mammalian cells.

Keywords

Chinese hamster ovary cells Human DNA topoisomerase I gene (TOP1) Matrix attachment region (MAR) Transgene expression 

Abbreviations

EGFP

Enhanced green fluorescence protein

MAR

Matrix attachment region

MFI

Mean fluorescence intensity

qPCR

Real-time quantitative PCR

TOP1

Human DNA topoisomerase I gene

Notes

Acknowledgements

This work was supported by the Grants from the National Natural Science Foundation of China (No. 81673337), the Key Science and Technology Project of Henan (Grant Nos. 192102310149, 182102310194), the Key Scientific Research Projects in Universities of Henan Province (No. 18A350008), the National Natural Science Foundation of China (Grant No. U1804168), the Plan of Scientific and Technological Innovation Team for University, Henan Province, China (Grant No. 18IRTSTHN027) and the Industry-University-Research Project of Xinxiang Medical University (Grant No. 2017CXY-2-1).

Compliance with ethical standards

Conflict of interests

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Yan-Long Jia
    • 1
    • 2
  • Xiao Guo
    • 1
    • 2
  • Xi-Cheng Wang
    • 1
    • 2
  • Tian-Yun Wang
    • 2
    • 3
    Email author
  1. 1.School of PharmacyXinxiang Medical UniversityXinxiangChina
  2. 2.International Joint Research Laboratory for Recombiant Pharmaceutical Protein Expression System of HenanXinxiang Medical UniversityXinxiangChina
  3. 3.School of Basic MedicineXinxiang Medical UniversityXinxiangChina

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