title: Engineering Sialic Acid Synthesis Ability in Insect Cells.
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.
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: Karthik
authors.initials: K
authors.lastname: Viswanathan
authors.forename: Someet
authors.initials: S
authors.lastname: Narang
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Betenbaugh
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods; D006023:Glycoproteins / Q000235:genetics; D007313:Insecta / Q000235:genetics*; D019158:N-Acetylneuraminic Acid / Q000096:biosynthesis*; D011134:Polysaccharides / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D018411:Spodoptera / Q000235:genetics
chemical_list: D006023:Glycoproteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D019158:N-Acetylneuraminic Acid