Protocol for CRISPR-Cas9 modification of glycosylation in 3D organotypic skin models
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- Protocol for CRISPR-Cas9 modification of glycosylation in 3D organotypic skin models
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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).
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 100668 |
| Tidsskrift | STAR Protocols |
| Vol/bind | 2 |
| Udgave nummer | 3 |
| ISSN | 2666-1667 |
| DOI | |
| Status | Udgivet - 2021 |
Bibliografisk note
Funding Information:
This work was supported by the European Commission (GlycoSkin H2020-ERC), European Commission (Imgene H2O20), European Commission (Remodel), Lundbeck Foundation , the Danish Research Councils (Sapere Aude Research Leader grant to H.H.W.), Danish National Research Foundation ( DNRF107 ), the Friis Foundation , the Michelsen Foundation , A.P. Møller og Hustru Chastine Mc-Kinney Møllers Fond til Almene Formaal , Danish Strategic Research Council , Lundbeck Foundation , and the program of excellence from the University of Copenhagen ( CDO2016 ). We thank Karin Uch Hansen, Birgit Poulsen, Karen Biré, and Louise Rosgaard Duus for the excellent technical assistance and Johan H. Wandall for the music for the video. We acknowledge the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen. We would like to thank James G. Rheinwald, Harvard Institute of Medicine, for providing the N/TERT-1 cells.
Funding Information:
This work was supported by the European Commission (GlycoSkin H2020-ERC), European Commission (Imgene H2O20), European Commission (Remodel), Lundbeck Foundation, the Danish Research Councils (Sapere Aude Research Leader grant to H.H.W.), Danish National Research Foundation (DNRF107), the Friis Foundation, the Michelsen Foundation, A.P. M?ller og Hustru Chastine Mc-Kinney M?llers Fond til Almene Formaal, Danish Strategic Research Council, Lundbeck Foundation, and the program of excellence from the University of Copenhagen (CDO2016). We thank Karin Uch Hansen, Birgit Poulsen, Karen Bir?, and Louise Rosgaard Duus for the excellent technical assistance and Johan H. Wandall for the music for the video. We acknowledge the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen. We would like to thank James G. Rheinwald, Harvard Institute of Medicine, for providing the N/TERT-1 cells. I.N.M. and S.D. wrote the manuscript, and all authors edited and approved the final version. The authors declare no competing interests.
Publisher Copyright:
© 2021 The Authors
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