RNA Interference mediated knockdown of genes in order to increase protein production using the baculovirus expression system
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KeywordsGreen Fluorescent Protein Recombinant Baculovirus Baculovirus Expression System Reduce Cell Death Infected Cell Culture
The baculovirus expression system has proven to be a robust and versatile system for recombinant protein production in insect cells. A wide range of promoters is available for the facile expression of transgenes, and yields of up to 50% of total protein have been reported . However, in many cases yield is decreased as a result of proteases and host cell apoptosis . Past efforts to overcome this problem include co-expressing chaperone proteins to assist with folding , anti-apoptotic proteins to reduce cell death, or adding chemical protease inhibitors to the culture media . However, these methods may have non-specific effects, prove too costly to be practical, or impose an undue metabolic burden on an already stressed cell. An alternative approach to increasing protein production is through the application of RNA interference (RNAi) to knockdown viral and host genes responsible for decreasing the yield of recombinant protein. Potential targets include proteases, cell-death proteins, and cell cycle regulators. By altering the metabolic landscape of cells prior to the introduction of the baculovirus, protein production can be improved.
Overall, our results support the application of RNAi as a metabolic engineering tool, specifically for enhancing protein productivity in the baculovirus expression system.
This article is published under license to BioMed Central Ltd.