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Protein expression/secretion boost by a novel unique 21-mer cis-regulatory motif (Exin21) via mRNA stabilization
Zhu, Yuanjun Saribas, A. Sami Liu, Jinbiao Lin, Yuan Zhao, Ruotong Guo, Qian Ting, Julia Wei, Zhengyu Ellis, Aidan
Zhu, Yuanjun
Saribas, A. Sami
Liu, Jinbiao
Lin, Yuan
Zhao, Ruotong
Guo, Qian
Ting, Julia
Wei, Zhengyu
Ellis, Aidan
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Journal article
Date
2023-04-05
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Pathology and Laboratory Medicine
Medical Genetics & Molecular Biochemistry
Medical Genetics & Molecular Biochemistry
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https://doi.org/10.1016/j.ymthe.2023.02.012
Abstract
Boosting protein production is invaluable in both industrial and academic applications. We discovered a novel expression-increasing 21-mer cis-regulatory motif (Exin21) that inserts between SARS-CoV-2 envelope (E) protein-encoding sequence and luciferase reporter gene. This unique Exin21 (CAACCGCGGTTCGCGGCCGCT), encoding a heptapeptide (QPRFAAA, designated as Qα), significantly (34-fold on average) boosted E production. Both synonymous and nonsynonymous mutations within Exin21 diminished its boosting capability, indicating the exclusive composition and order of 21 nucleotides. Further investigations demonstrated that Exin21/Qα addition could boost the production of multiple SARS-CoV-2 structural proteins (S, M, and N) and accessory proteins (NSP2, NSP16, and ORF3), and host cellular gene products such as IL-2, IFN-γ, ACE2, and NIBP. Exin21/Qα enhanced the packaging yield of S-containing pseudoviruses and standard lentivirus. Exin21/Qα addition on the heavy and light chains of human anti-SARS-CoV monoclonal antibody robustly increased antibody production. The extent of such boosting varied with protein types, cellular density/function, transfection efficiency, reporter dosage, secretion signaling, and 2A-mediated auto-cleaving efficiency. Mechanistically, Exin21/Qα increased mRNA synthesis/stability, and facilitated protein expression and secretion. These findings indicate that Exin21/Qα has the potential to be used as a universal booster for protein production, which is of importance for biomedicine research and development of bioproducts, drugs, and vaccines.
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Citation
Yuanjun Zhu, A. Sami Saribas, Jinbiao Liu, Yuan Lin, Brittany Bodnar, Ruotong Zhao, Qian Guo, Julia Ting, Zhengyu Wei, Aidan Ellis, Fang Li, Xu Wang, Xiaofeng Yang, Hong Wang, Wen-Zhe Ho, Ling Yang, Wenhui Hu, Protein expression/secretion boost by a novel unique 21-mer cis-regulatory motif (Exin21) via mRNA stabilization, Molecular Therapy, Volume 31, Issue 4, 2023, Pages 1136-1158, ISSN 1525-0016, https://doi.org/10.1016/j.ymthe.2023.02.012. (https://www.sciencedirect.com/science/article/pii/S1525001623000758)
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Cell Press
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Molecular Therapy, Vol. 31, Iss. 4
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