The GAGOme: a cell-based library of displayed glycosaminoglycans
The GAGOme: a cell-based library of displayed glycosaminoglycans
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DOI：
10.1038/s41592-018-0086-z
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发表时间：
2018-11-01
期刊：
影响因子：
48
通讯作者：
Yang, Zhang
中科院分区：
文献类型：
--
作者：
Chen, Yen-Hsi;Narimatsu, Yoshiki;Yang, Zhang
糖胺聚糖（Glycosaminoglycans，GAG）是人体正常生理和疾病中必需的多糖。然而，对特定GAG结构对特定生物学功能的贡献的理解是有限的，主要是因为GAG本身之间的巨大结构异质性，以及GAG的结构表征和化学合成的技术限制。在这里，我们描述了一种基于细胞的方法来产生和展示具有广泛修饰库的不同GAG，我们将该文库称为GAGOme。通过使用精确的基因编辑，我们对大量中国仓鼠卵巢细胞进行了基因敲除或敲入，以产生差异显示不同GAG特征的等基因细胞系库，这些细胞编码参与GAG生物合成的大多数酶。我们表明，该库可用于基于细胞的结合试验，具有不同GAG结构的蛋白聚糖的重组表达，以及在可用于组装GAG聚糖微阵列的代谢引物上产生不同的GAG链。
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.