Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, University of Copenhagen, Copenhagen, Denmark.
Department of Oral Pathology, School of Dentistry, University of Copenhagen, Copenhagen, Denmark.
STAR Protoc. 2021 Jul 10;2(3):100668. doi: 10.1016/j.xpro.2021.100668. eCollection 2021 Sep 17.
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).
糖基化是生物体内最常见的蛋白质修饰之一，在动物组织发育和动态平衡中具有重要的调节作用。本文提供了一种使用 CRISPR-Cas9 基因工程人角质形成细胞（N/TERT-1）生成 3D 器官型皮肤模型的方案，用于研究糖在上皮组织形成中的作用。该策略也适用于其他基因靶标和器官型组织模型。细胞培养物的小心处理对于类器官的成功形成至关重要。有关该方案使用和执行的完整详细信息，请参阅 Dabelsteen 等人（2020 年）。
