Optimization protein productivity of human interleukin-2 through codon usage, gene copy number and intracellular tRNA concentration in CHO cells

Kua Chun Ou, Chih Yang Wang, Kuan Ting Liu, Yi Ling Chen, Yi Chen Chen, Ming Derg Lai, Meng Chi Yen

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Transfer RNA (tRNA) abundance is one of the critical factors for the enhancement of protein productivity in prokaryotic and eukaryotic hosts. Gene copy number of tRNA and tRNA codon usage bias are generally used to match tRNA abundance of protein-expressing hosts and to optimize the codons of recombinant proteins. Because sufficient concentration of intracellular tRNA and optimized codons of recombinant proteins enhanced translation efficiency, we hypothesized that sufficient supplement of host's tRNA improved protein productivity in mammalian cells. First, the small tRNA sequencing results of CHO-K1 cells showed moderate positive correlation with gene copy number and codon usage bias. Modification of human interleukin-2 (IL-2) through codons with high gene copy number and high codon usage bias (IL-2 HH, modified on Leu, Thr, Glu) significantly increased protein productivity in CHO-K1 cells. In contrast, modification through codons with relatively high gene copy number and low codon usage bias (IL-2 HL, modified on Ala, Thr, Val), or relatively low gene copy number and low codon usage bias (IL-2 LH, modified on Ala, Thr, Val) did not increase IL-2 productivity significantly. Furthermore, supplement of the alanine tRNA or threonine tRNA increased IL-2 productivity of IL-2 HL. In summary, we revealed a potential strategy to enhance productivity of recombinant proteins, which may be applied in production of protein drug or design of DNA vaccine.

Original languageEnglish
Pages (from-to)347-352
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume454
Issue number2
DOIs
Publication statusPublished - Nov 14 2014
Externally publishedYes

Fingerprint

Gene Dosage
CHO Cells
Transfer RNA
Codon
Genes
Productivity
Interleukin-2
Recombinant Proteins
Proteins
RNA Sequence Analysis
human IL2 protein
DNA Vaccines
Drug Design
Protein Biosynthesis
Threonine
Alanine
Cells

Keywords

  • Codon usage bias
  • DNA vaccine
  • Gene copy number
  • Interleukin-2
  • tRNA abundance

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Optimization protein productivity of human interleukin-2 through codon usage, gene copy number and intracellular tRNA concentration in CHO cells. / Ou, Kua Chun; Wang, Chih Yang; Liu, Kuan Ting; Chen, Yi Ling; Chen, Yi Chen; Lai, Ming Derg; Yen, Meng Chi.

In: Biochemical and Biophysical Research Communications, Vol. 454, No. 2, 14.11.2014, p. 347-352.

Research output: Contribution to journalArticle

Ou, Kua Chun ; Wang, Chih Yang ; Liu, Kuan Ting ; Chen, Yi Ling ; Chen, Yi Chen ; Lai, Ming Derg ; Yen, Meng Chi. / Optimization protein productivity of human interleukin-2 through codon usage, gene copy number and intracellular tRNA concentration in CHO cells. In: Biochemical and Biophysical Research Communications. 2014 ; Vol. 454, No. 2. pp. 347-352.
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