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Methods Enzymol. 2010;480:477-93. doi: 10.1016/S0076-6879(10)80021-7.

High-throughput RNAi screening for N-glycosylation dependent loci in Caenorhabditis elegans.

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National Institute for Bioprocessing Research and Training, Dublin-Oxford Glycobiology Group, Conway Institute for Biomolecular and Biomedical Sciences, University College Dublin, Belfield, Dublin, Ireland.


The attachment of oligosaccharides to the amide nitrogen of asparagine side chains on proteins is a fundamental process occurring in all metazoans. This process, known as N-glycosylation, is complex and is achieved by the precise interactions of various cellular components. The initial stage of N-glycan biosynthesis is preserved among eukaryotes, and defective enzymes or components in this pathway cause congenital disorders of glycosylation type I (CDG-I) in humans. This disease is rare but exceedingly life-threatening with no known cure. Paramount to CDG treatment and care is understanding the mechanisms of N-glycosylation and factors that influence the pathology of the disease, both of which are not completely known. Here we outline a novel technique to model a CDG-I-like condition and identify genes that are vital for healthy glycosylation in Caenorhabditis elegans. C. elegans is a well-established model for understanding the complexity of glycosylation in development and disease. Although C. elegans N-glycan structures are dissimilar to that observed in higher eukaryotes, they contain over 150 gene homologs that are directly involved in glycosylation. Moreover, the annotated genome of C. elegans, its susceptibility to genetic silencing and its recognizable phenotypes, is a suitable model to dissect the complex phenomenon of glycosylation and identify genes that are required for N-glycan biosynthesis.

[Indexed for MEDLINE]

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