Efficient Human Cell Coexpression System and Its Application to the Production of Multiple Coronavirus Antigens

Chonsaeng Kim, You Kyeong Jeong, Jihyeon Yu, Hye Jin Shin, Keun Bon Ku, Hyung Jin Cha, Jun Hee Han, Sung Ah Hong, Bum Tae Kim, Seong Jun Kim, Jae Sung Woo, Sangsu Bae

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)


Coproduction of multiple proteins at high levels in a single human cell line would be extremely useful for basic research and medical applications. Here, a novel strategy for the stable expression of multiple proteins by integrating the genes into defined transcriptional hotspots in the human genome is presented. As a proof-of-concept, it is shown that EYFP is expressed at similar levels from hotspots and that the EYFP expression increases proportionally with the copy number. It is confirmed that three different fluorescent proteins, encoded by genes integrated at different loci, can be coexpressed at high levels. Further, a stable cell line is generated, producing antigens from different human coronaviruses: MERS-CoV and HCoV-OC43. Antibodies raised against these antigens, which contain human N-glycosylation, show neutralizing activities against both viruses, suggesting that the coexpression system provides a quick and predictable way to produce multiple coronavirus antigens, such as the recent 2019 novel human coronavirus.

Original languageEnglish
Article number2000154
JournalAdvanced Biology
Issue number4
Publication statusPublished - 2021 Apr


  • CRISPR-Cas9
  • coronaviruses
  • protein expression
  • targeted knock-in
  • vaccines

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biomedical Engineering
  • Biomaterials
  • Medicine(all)


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