Improvement of protein production by engineering a novel antiapoptotic baculovirus vector to suppress the expression of Sf‐caspase‐1 and Tn‐caspase‐1. Issue 8 (15th May 2021)

• Record Type: Journal Article
• Authors: Zhang, Xiaoyue; Zhao, Kaixia; Lan, Lan; Shi, Na; Nan, Hao; Shi, Yanan; Xu, Xiaodong; Chen, Hongying
• Is Part Of: Biotechnology and bioengineering. Volume 118:Issue 8(2021)
• Page Start: 2977
• Page End: 2989
• Publication Date: 2021-05-15
• Subjects: apoptosis -- baculovirus expression vector system -- Sf‐caspase‐1 -- shRNA -- Tn‐caspase‐1; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.27807
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: The baculovirus expression vector system (BEVS) is an attractive manufacturing platform for recombinant protein production in insect cells. However, baculovirus infection commonly induces host apoptosis in 3–4 days which would subsequently terminate the protein expression. Previous studies have proved that protein production by BEVS can be elevated in apoptosis‐suppressed insect cells. We also developed a baculovirus vector in our previous report to inhibit the apoptosis and improve protein production in Sf 9 cells. In this study, we designed five short hairpin RNA (shRNA) expression cassettes targeting a conserved region in Spodoptera frugiperda caspase‐1 ( Sf‐caspase‐1 ) and Trichoplusia ni caspase‐1 ( Tn‐caspase‐1 ), and found that introduction of C to T mutations within the stem region of the expression cassette was beneficial for the heterologous protein expression. One of the improved shRNA expression cassettes was knocked into a bacmid with the deletion of several nonessential genes. The novel baculovirus vector demonstrated the ability to suppress cell apoptosis in both Sf 9 and High Five cells, and exhibited superior recombinant protein productivity of intracellularly expressed GFP and firefly luciferase and secreted glycoprotein OD‐Fc. The antiapoptotic baculovirus vector developed in this study could serve as a useful tool for the protein production in scientific research and pharmaceutical industries. Abstract : A novel baculovirus vector, which carries an shRNA expression cassette targeting a conserved sequence of Sf‐caspase‐1 and Tn‐caspase‐1, was developed in this study. Using this vector, baculovirus‐induced cell apoptosis was inhibited, and the production of recombinant proteins were significantly increased in both Sf9 and High Five cells.