Essential Functions of Glycans in Human Epithelia Dissected by a CRISPR-Cas9-Engineered Human Organotypic Skin Model

Sally Dabelsteen, Emil M H Pallesen, Irina N Marinova, Mathias I Nielsen, Maria Adamopoulou, Troels B Rømer, Asha Levann, Mikkel M Andersen, Zilu Ye, David Thein, Eric P Bennett, Christian Büll, Sam J Moons, Thomas Boltje, Henrik Clausen, Sergey Y Vakhrushev, Ieva Bagdonaite, Hans H Wandall


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.

Original languageEnglish
JournalDevelopmental Cell
Issue number5
Pages (from-to)669-684.e7
Publication statusPublished - 14 Sep 2020


  • CRISPR-Cas Systems/genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats/genetics
  • Epithelium/physiology
  • Gene Library
  • Glycoproteins/genetics
  • Glycosylation
  • Humans
  • Polysaccharides/genetics
  • Skin/metabolism


Dive into the research topics of 'Essential Functions of Glycans in Human Epithelia Dissected by a CRISPR-Cas9-Engineered Human Organotypic Skin Model'. Together they form a unique fingerprint.

Cite this