Protocol for CRISPR-Cas9 modification of glycosylation in 3D organotypic skin models

Glycosylation is one of the most common protein modifications in living organisms and has important regulatory roles in animal tissue development and homeostasis. Here, we present a protocol for generation of 3D organotypic skin models using CRISPR-Cas9 genetically engineered human keratinocytes (N/TERT-1) to study the role of glycans in epithelial tissue formation. This strategy is also applicable to other gene targets and organotypic tissue models. Careful handling of the cell cultures is critical for the successful formation of the organoids.For complete details on the use and execution of this protocol, please refer to Dabelsteen et al. (2020).

Automated summary

The study introduces a protocol for generating 3D organotypic skin models using CRISPR-Cas9 genetically engineered human keratinocytes to study the role of glycans in epithelial tissue formation. This strategy is also applicable to other gene targets and tissue models, emphasizing the importance of careful handling of cell cultures for successful organoid formation. For full details, please refer to Dabelsteen et al. (2020).