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Stabilizing and Anti-Repressor Elements Effectively Increases Transgene Expression in Transfected CHO Cells.

Qin Li1,2, Rui-Fang Yan1, Yong-Xiao Yang2

  • 1School of Basic Medicine, Xinxiang Medical University, Xinxiang, China.

Frontiers in Bioengineering and Biotechnology
|June 20, 2022
PubMed
Summary

Stabilizing and anti-repressor (SAR) elements enhance recombinant therapeutic protein (RTP) production in Chinese hamster ovary (CHO) cells. SAR7 and SAR40 elements significantly boosted and sustained long-term transgene expression, offering a promising strategy for increasing RTP yields.

Keywords:
Chinese hamster ovary cellepigenetic regulatory elementsgreen fluorescent proteinstabilizing and anti-repressor elementstransgene expression

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Cell Biology

Background:

  • Chinese hamster ovary (CHO) cells are the primary host for recombinant therapeutic protein (RTP) production.
  • Increasing RTP yields is crucial for meeting market demand and reducing manufacturing costs.

Purpose of the Study:

  • To investigate the efficacy of human stabilizing and anti-repressor (SAR) elements in enhancing transgene expression in CHO cells.
  • To evaluate the long-term expression stability and impact on gene copy and mRNA levels.

Main Methods:

  • Cloning of human SAR7, SAR40, and SAR44 elements into eukaryotic vectors upstream of the promoter.
  • Transfection into CHO cells and screening under G418 selection.
  • Analysis of enhanced green fluorescent protein (eGFP) expression via flow cytometry.
  • Quantification of gene copy and mRNA expression levels using quantitative real-time PCR.
  • Assessment of SAR element effects on adalimumab expression.

Main Results:

  • Transgene expression levels were significantly higher in SAR-containing vectors compared to the control.
  • SAR7 and SAR40 elements demonstrated a notable increase and sustained long-term transgene expression.
  • Enhanced transgene expression correlated with increased gene copy numbers and mRNA expression levels.
  • The impact of SAR elements was also observed for adalimumab expression.

Conclusions:

  • Human SAR elements can effectively enhance transgene expression in CHO cells.
  • SAR7 and SAR40 promote stable, long-term transgene expression, crucial for efficient RTP production.
  • This strategy holds potential for optimizing recombinant protein manufacturing in CHO cell lines.