Abstract
Insect cells lack the ability to synthesize the sialic acid donor molecule CMP-sialic acid or its precursor, sialic acid. In this chapter, we describe a method to engineer CMP-sialic acid synthesis capability into Spodoptera frugiperda (Sf9) cells, a prototypical insect cell line, by recombinant expression of sialic acid synthesis pathway genes using baculovirus technology. Co-expression of a sialuria mutant UDP-GlcNAc- 2-epimerase/ManNAc kinase (EKR263L), wild-type sialic acid 9-phosphate synthase (SAS), and wild-type CMP-sialic acid synthetase (CSAS) in the presence of GlcNAc leads to synthesis of CMP-sialic acids synthesis to support sialylation of N-glycans on glycoproteins.
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CITATION STYLE
Viswanathan, K., Narang, S., & Betenbaugh, M. J. (2015). Engineering sialic acid synthesis ability in insect cells. In Glyco-Engineering: Methods and Protocols (pp. 171–178). Springer New York. https://doi.org/10.1007/978-1-4939-2760-9_12
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