Comparison of three transposons for the generation of highly productive recombinant CHO cell pools and cell lines

Biotechnol Bioeng. 2016 Jun;113(6):1234-43. doi: 10.1002/bit.25888. Epub 2015 Dec 9.


Several naturally occurring vertebrate transposable elements have been genetically modified to enable the transposition of recombinant genes in mammalian cells. We compared three transposons-piggyBac, Tol2, and Sleeping Beauty-for their ability to generate cell pools (polyclonal cultures of recombinant cells) and clonal cell lines for the large-scale production of recombinant proteins using Chinese hamster ovary cells (CHO-DG44) as the host. Transfection with each of the dual-vector transposon systems resulted in cell pools with volumetric yields of tumor necrosis factor receptor-Fc fusion protein (TNFR:Fc) that were about ninefold higher than those from cell pools generated by conventional plasmid transfection. On average, the cell pools had 10-12 integrated copies of the transgene per cell. In the absence of selection, the volumetric productivity of the cell pools decreased by 50% over a 2-month cultivation period and then remained constant. The average volumetric TNFR:Fc productivity of clonal cell lines recovered from cell pools was about 25 times higher than that of cell lines generated by conventional transfection. In 14-day fed-batch cultures, TNFR:Fc levels up to 900 mg/L were obtained from polyclonal cell pools and up to 1.5 g/L from clonal cell lines using any of the three transposons. Biotechnol. Bioeng. 2016;113: 1234-1243. © 2015 Wiley Periodicals, Inc.

Keywords: Sleeping Beauty; Tol2; cell pools; piggyBac; recombinant protein; transposon.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Animals
  • Batch Cell Culture Techniques / methods*
  • CHO Cells
  • Cricetulus / genetics
  • Cricetulus / metabolism
  • DNA Transposable Elements / genetics*
  • Escherichia coli Proteins / genetics
  • Genetic Enhancement / methods*
  • Nerve Tissue Proteins / genetics
  • Protein Engineering / methods*
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics*
  • Transposases / genetics


  • DNA Transposable Elements
  • Escherichia coli Proteins
  • Nerve Tissue Proteins
  • PGBD1 protein, human
  • Recombinant Proteins
  • tolA protein, E coli
  • Transposases
  • sleeping beauty transposase, human