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The GAGOme : a cell-based library of displayed glycosaminoglycans

Chen, Yen Hsi; Narimatsu, Yoshiki; Clausen, Thomas M.; Gomes, Catarina; Karlsson, Richard; Steentoft, Catharina; Spliid, Charlotte B.; Gustavsson, Tobias LU ; Salanti, Ali and Persson, Andrea LU , et al. (2018) In Nature Methods 15(11). p.881-888
Abstract

Glycosaminoglycans (GAGs) are essential polysaccharides in normal physiology and disease. However, understanding of the contribution of specific GAG structures to specific biological functions is limited, largely because of the great structural heterogeneity among GAGs themselves, as well as technical limitations in the structural characterization and chemical synthesis of GAGs. Here we describe a cell-based method to produce and display distinct GAGs with a broad repertoire of modifications, a library we refer to as the GAGOme. By using precise gene editing, we engineered a large panel of Chinese hamster ovary cells with knockout or knock-in of the genes encoding most of the enzymes involved in GAG biosynthesis, to generate a library... (More)

Glycosaminoglycans (GAGs) are essential polysaccharides in normal physiology and disease. However, understanding of the contribution of specific GAG structures to specific biological functions is limited, largely because of the great structural heterogeneity among GAGs themselves, as well as technical limitations in the structural characterization and chemical synthesis of GAGs. Here we describe a cell-based method to produce and display distinct GAGs with a broad repertoire of modifications, a library we refer to as the GAGOme. By using precise gene editing, we engineered a large panel of Chinese hamster ovary cells with knockout or knock-in of the genes encoding most of the enzymes involved in GAG biosynthesis, to generate a library of isogenic cell lines that differentially display distinct GAG features. We show that this library can be used for cell-based binding assays, recombinant expression of proteoglycans with distinct GAG structures, and production of distinct GAG chains on metabolic primers that may be used for the assembly of GAG glycan microarrays.

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published
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Nature Methods
volume
15
issue
11
pages
881 - 888
publisher
Nature Publishing Group
external identifiers
  • scopus:85052313120
ISSN
1548-7091
DOI
10.1038/s41592-018-0086-z
language
English
LU publication?
yes
id
4888f4e7-cab2-48ee-880e-691cd00b7fbb
date added to LUP
2018-10-05 10:54:20
date last changed
2019-03-19 04:00:15
@article{4888f4e7-cab2-48ee-880e-691cd00b7fbb,
  abstract     = {<p>Glycosaminoglycans (GAGs) are essential polysaccharides in normal physiology and disease. However, understanding of the contribution of specific GAG structures to specific biological functions is limited, largely because of the great structural heterogeneity among GAGs themselves, as well as technical limitations in the structural characterization and chemical synthesis of GAGs. Here we describe a cell-based method to produce and display distinct GAGs with a broad repertoire of modifications, a library we refer to as the GAGOme. By using precise gene editing, we engineered a large panel of Chinese hamster ovary cells with knockout or knock-in of the genes encoding most of the enzymes involved in GAG biosynthesis, to generate a library of isogenic cell lines that differentially display distinct GAG features. We show that this library can be used for cell-based binding assays, recombinant expression of proteoglycans with distinct GAG structures, and production of distinct GAG chains on metabolic primers that may be used for the assembly of GAG glycan microarrays.</p>},
  author       = {Chen, Yen Hsi and Narimatsu, Yoshiki and Clausen, Thomas M. and Gomes, Catarina and Karlsson, Richard and Steentoft, Catharina and Spliid, Charlotte B. and Gustavsson, Tobias and Salanti, Ali and Persson, Andrea and Malmström, Anders and Willén, Daniel and Ellervik, Ulf and Bennett, Eric P. and Mao, Yang and Clausen, Henrik and Yang, Zhang},
  issn         = {1548-7091},
  language     = {eng},
  number       = {11},
  pages        = {881--888},
  publisher    = {Nature Publishing Group},
  series       = {Nature Methods},
  title        = {The GAGOme : a cell-based library of displayed glycosaminoglycans},
  url          = {http://dx.doi.org/10.1038/s41592-018-0086-z},
  volume       = {15},
  year         = {2018},
}