Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Jan;47(1):469-475.
doi: 10.1007/s11033-019-05153-3. Epub 2019 Oct 28.

Effects of different 2A peptides on transgene expression mediated by tricistronic vectors in transfected CHO cells

Yan-Mei Li  1   2 , Meng Wang  1   2 , Tian-Yun Wang  3   4 , Yong-Ge Wei  1   2 , Xiao Guo  1   2 , Chun-Liu Mi  1   2 , Chun-Peng Zhao  1   2 , Xiang-Xiang Cao  1   2 , Yuan-Yuan Dou  1   2
Affiliations

Effects of different 2A peptides on transgene expression mediated by tricistronic vectors in transfected CHO cells

Yan-Mei Li et al. Mol Biol Rep. 2020 Jan.

Abstract

Multicistronic vectors can increase transgene expression and decrease the imbalance of gene expression in the Chinese hamster ovary (CHO) cell expression system. Small, self-cleaving 2A peptides have a high cleavage efficiency and are essential for constructing high-expression multicistronic vectors. In this study, we investigated the effects of two different 2A peptides on transgene expression in CHO cells via their mediating action on tricistronic vectors. The enhanced green fluorescent protein (eGFP) and red fluorescent protein (RFP) genes were linked by the porcine teschovirus-1 (P2A) and Thosea asigna virus (T2A) peptides in a multicistronic vector. We transfected CHO cells with these vectors and screened for the presence of blasticidin-resistant colonies. Flow cytometry and real-time quantitative PCR (qPCR) were used to detect the expression levels of eGFP and RFP and the copy numbers of stably transfected cells. The results showed that P2A could enhance eGFP and RFP expression by 1.48- and 1.47-fold, respectively, compared to T2A. The expression levels of the genes were not proportional to their copy numbers. In conclusion, we found that P2A can effectively drive transgene expression in CHO cells and a potent 2A peptide can be used for recombinant protein production in the CHO cell system.

Keywords: 2A peptides; CHO cells; Transgene expression; Tricistronic.

PubMed Disclaimer

References

    1. PLoS One. 2011;6(4):e18556 - PubMed
    1. Nat Commun. 2015 Aug 27;6:8072 - PubMed
    1. Biochemistry (Mosc). 2016 May;81(5):521-9 - PubMed
    1. J Gene Med. 2000 Jul-Aug;2(4):243-9 - PubMed
    1. Prep Biochem Biotechnol. 2018 Feb 7;48(2):160-164 - PubMed

MeSH terms

LinkOut - more resources