An anti-apoptotic baculovirus expression vector

By introducing small RNA expression cassettes targeting Sf-caspase-1 and Tn-caspase-1 into baculovirus vectors, the problem of low recombinant protein yield caused by insect cell apoptosis was solved, enabling efficient expression of exogenous genes in Sf and Tn cell lines and advancing the industrial application of anti-apoptotic vectors.

CN116144653BActive Publication Date: 2026-04-14SHAANXI BACMID BIOTECHNOLOGY CO LTD
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Patent Information

Application Number
CN202211099000.2
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2018-06-21
Publication Date
2026-04-14
Estimated Expiration
2038-06-21

AI Technical Summary

Technical Problem

In existing baculovirus expression systems, insect cells undergo apoptosis shortly after infection, resulting in limited recombinant protein production. In particular, the lack of simultaneously effective anti-apoptotic vectors in Sf and Tn cell lines limits their industrial application.

Method used

We designed and cloned a small RNA expression cassette targeting the shared sequences of Sf-caspase-1 and Tn-caspase-1 into the genome of a baculovirus vector. The cassette contained the human U6 promoter, a 21nt guide strand complement sequence, a 9nt loop sequence, and a transcription termination signal, which reduced the tendency of the DNA strand to form hairpin structures and constructed an anti-apoptotic recombinant baculovirus vector.

Benefits of technology

It significantly improved the expression level of exogenous genes and protein production in Sf9 and High Five cells, increased luciferase activity by about 1 time, and prolonged the survival time of infected cells, making it suitable for the industrial production of protein drugs and vaccines.

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Abstract

The application relates to an anti-apoptosis baculovirus expression vector, which realizes a broad-spectrum anti-apoptosis effect by expressing siRNA targeting a Sf-caspase-1 and Tn-caspase-1 common sequence of Spodoptera frugiperda and Trichoplusia ni insect cells through the vector. The baculovirus vector contains a specific DNA sequence, the DNA sequence contains an siRNA sequence targeting the Sf-caspase-1 and Tn-caspase-1 common sequence transcribed by an RNA polymerase III promoter; the recombinant virus of the vector can express double-stranded small RNA in a host cell, silences the caspase-1 encoded by the host cell through an RNA interference pathway, thereby inhibiting the apoptosis of the host cell, and significantly improving the expression level of an exogenous protein. The application can be used for industrialized production of protein preparations and vaccines.
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