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Two human MARs effectively increase transgene expression in transfected CHO cells.

Qin Li1,2, Chun-Peng Zhao1,2, Yan Lin1,2

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Journal of Cellular and Molecular Medicine
|November 20, 2018
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Two new human matrix attachment regions (MARs), MAR-3 and MAR-7, significantly enhance transgene expression in Chinese hamster ovary (CHO) cells. These MAR elements may boost expression by increasing transgene copy numbers.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genetics

Background:

  • Matrix attachment regions (MARs) are DNA elements known to influence transgene expression.
  • The precise mechanisms by which MARs enhance gene expression and their functional improvements require further investigation.
  • Identifying novel and more functional MARs is crucial for optimizing gene expression systems.

Purpose of the Study:

  • To screen and characterize novel, highly functional matrix attachment regions (MARs) from the human genome.
  • To evaluate the efficacy of two identified human MAR elements (MAR-3 and MAR-7) in enhancing transgene expression in a Chinese hamster ovary (CHO) cell system.
  • To elucidate the potential mechanisms underlying MAR-mediated transgene expression enhancement.

Main Methods:

  • Cloning of human MAR-3 and MAR-7 elements downstream of the polyA site in a eukaryotic expression vector.
  • Transfection of CHO cells with MAR-containing and control vectors, followed by selection using G418.
  • Quantification of enhanced green fluorescent protein (eGFP) expression levels and stability via flow cytometry.
  • Assessment of transgene copy number and mRNA expression levels using quantitative real-time PCR.

Main Results:

  • MAR-containing vectors demonstrated significantly higher eGFP expression levels compared to control vectors in both transient and stable transfections.
  • The MAR-7 element exhibited a greater enhancing effect on eGFP expression than the MAR-3 element.
  • MAR elements were found to significantly increase the copy number of the eGFP transgene and its corresponding mRNA expression levels.

Conclusions:

  • Human MAR-3 and MAR-7 elements effectively promote transgene expression in transfected CHO cells.
  • The mechanism of MAR-mediated enhancement appears to be associated with an increase in transgene copy number.
  • These findings provide valuable MAR elements for improving transgene expression in biotechnological applications.