The glycome undergoes characteristic changes during histogenesis and organogenesis, but our understanding of the importance of select glycan structures for tissue formation and homeostasis is incomplete. Here, we present a human organotypic platform that allows genetic dissection of cellular glycosylation capacities and systematic interrogation of the roles of distinct glycan types in tissue formation. We used CRISPR-Cas9 gene targeting to generate a library of 3D organotypic skin tissues that selectively differ in their capacity to produce glycan structures on the main types of N- and O-linked glycoproteins and glycolipids. This tissue library revealed distinct changes in skin formation associated with a loss of features for all tested glycoconjugates. The organotypic skin model provides phenotypic cues for the distinct functions of glycoconjugates and serves as a unique resource for further genetic dissection and identification of the specific structural features involved. The strategy is also applicable to other organotypic tissue models.

糖蛋白在组织发生和器官发生过程中经历特征性变化，但是我们对选择的聚糖结构对于组织形成和体内平衡的重要性的理解尚不完全。在这里，我们介绍了一个人类器官型平台，该平台允许对细胞糖基化能力进行遗传解剖，并对不同聚糖类型在组织形成中的作用进行系统的研究。我们使用CRISPR-Cas9基因定位来生成3D器官型皮肤组织库，这些库在产生N型和O型连接糖蛋白和糖脂的主要类型上的聚糖结构的能力方面选择性地不同。该组织库显示出皮肤形成的明显变化与所有测试的糖缀合物的功能丧失有关。器官型皮肤模型为糖缀合物的独特功能提供了表型线索，并为进一步的遗传解剖和鉴定涉及的特定结构特征提供了独特的资源。该策略也适用于其他器官型组织模型。