Protein expression/secretion boost by a novel unique 21-mer cis-regulatory motif (Exin21) via mRNA stabilization.
Protein expression/secretion boost by a novel unique 21-mer cis-regulatory motif (Exin21) via mRNA stabilization.
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DOI：
10.1016/j.ymthe.2023.02.012
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发表时间：
2023-04-05
影响因子：
12.4
通讯作者：
Hu, Wenhui
中科院分区：
文献类型：
--
作者：
Zhu, Yuanjun;Saribas, A. Sami;Liu, Jinbiao;Lin, Yuan;Bodnar, Brittany;Zhao, Ruotong;Guo, Qian;Ting, Julia;Wei, Zhengyu;Ellis, Aidan;Li, Fang;Wang, Xu;Yang, Xiaofeng;Wang, Hong;Ho, Wen-Zhe;Yang, Ling;Hu, Wenhui
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. Hu and colleagues discovered a novel and unique cis-regulatory 21-mer motif that can remarkably boost protein production via increasing mRNA synthesis and stability. This breakthrough will open a new research avenue in the field of mRNA/protein homeostasis and facilitate versatile applications to the development of bioproducts, drugs, and vaccines.
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