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Improvement of protein production by engineering a novel antiapoptotic baculovirus vector to suppress the expression of Sf‐caspase‐1 and Tn‐caspase‐1
Author(s) -
Zhang Xiaoyue,
Zhao Kaixia,
Lan Lan,
Shi Na,
Nan Hao,
Shi Yanan,
Xu Xiaodong,
Chen Hongying
Publication year - 2021
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27807
Subject(s) - sf9 , small hairpin rna , expression cassette , biology , expression vector , spodoptera , rna interference , microbiology and biotechnology , apoptosis , heterologous , trichoplusia , transfection , green fluorescent protein , recombinant dna , caspase 3 , heterologous expression , cell culture , vector (molecular biology) , gene knockdown , rna , gene , programmed cell death , biochemistry , genetics , botany , larva , noctuidae
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.