Distance effect characteristic of the matrix attachment region increases recombinant protein expression in Chinese hamster ovary cells

Biotechnol Lett. 2020 Feb;42(2):187-196. doi: 10.1007/s10529-019-02775-2. Epub 2019 Nov 27.

Abstract

Objectives: Previously, we have found that the matrix attachment region (MAR) may confer a 'distance effect' on transgene expression. This work aims to systematically explore the increased transgene expression in transfected Chinese hamster ovary (CHO) cells due to the characteristics of MAR and its mechanism.

Results: Compared with the control vector, 500 and 1000 bp DNA distances between MAR and the cytomegalovirus promoter can increase transgene expression by 1.77- and 1.56-fold, respectively. Meanwhile, transgene expression was not affected when 2000 and 2500 bp spacer DNAs were inserted, but a declining trend was observed when a 1500 bp spacer DNA was inserted. The vector containing a 500 bp DNA distance significantly increased the expression of the enhanced green fluorescent protein, and this increase was not related to transgene copy numbers.

Conclusions: A short DNA distance-containing MAR confers high transgene expression level in transfected CHO cells, but a distance threshold does not exist in the vector system.

Keywords: Chinese hamster ovary cells; Distance effect; Matrix attachment region; Transgene expression.

MeSH terms

  • Animals
  • CHO Cells
  • Cloning, Molecular / methods*
  • Cricetinae
  • Cricetulus
  • Gene Expression
  • Matrix Attachment Regions
  • Promoter Regions, Genetic
  • Recombinant Proteins / metabolism*
  • Transfection
  • Transgenes*

Substances

  • Recombinant Proteins