Abstract
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 language | English |
|---|---|
| Article number | 2000154 |
| Journal | Advanced Biology |
| Volume | 5 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 2021 Apr |
Bibliographical note
Funding Information:The MERS-CoV was generously provided by Korea Centers for Disease Control and prevention. This research was supported by grants from the National Research Foundation of Korea (NRF) (No. 2018R1C1B6004447 to J.-S.W., No. 2021M3A9H3015389, and No. 2020M3A9I4036072 to S.B.), by the New Breeding Technologies Development Program (PJ01487401202001), and the Technology Innovation Program (No. 20000158) to S.B., and from the National Research Council of Science and Technology (NST) grant by the Ministry of Science and ICT (CRC-16-01-KRICT).
Funding Information:
The MERS‐CoV was generously provided by Korea Centers for Disease Control and prevention. This research was supported by grants from the National Research Foundation of Korea (NRF) (No. 2018R1C1B6004447 to J.‐S.W., No. 2021M3A9H3015389, and No. 2020M3A9I4036072 to S.B.), by the New Breeding Technologies Development Program (PJ01487401202001), and the Technology Innovation Program (No. 20000158) to S.B., and from the National Research Council of Science and Technology (NST) grant by the Ministry of Science and ICT (CRC‐16‐01‐KRICT).
Publisher Copyright:
© 2021 The Authors. Advanced Biology published by Wiley-VCH GmbH
Keywords
- CRISPR-Cas9
- coronaviruses
- protein expression
- targeted knock-in
- vaccines
ASJC Scopus subject areas
- General Biochemistry,Genetics and Molecular Biology
- Biomedical Engineering
- Biomaterials
