1. Protocol for CRISPR-Cas12a genome editing of protein tyrosine phosphatases in human pluripotent stem cells and functional β-like cell generation.
- Author
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Negueruela J, Vandenbempt V, Talamantes S, Ribeiro-Costa F, Nunes M, Dias A, Bansal M, and Gurzov EN
- Subjects
- Humans, Mice, Animals, Cell Differentiation genetics, Human Embryonic Stem Cells cytology, Human Embryonic Stem Cells metabolism, Gene Editing methods, CRISPR-Cas Systems genetics, Pluripotent Stem Cells cytology, Pluripotent Stem Cells metabolism, Insulin-Secreting Cells cytology, Insulin-Secreting Cells metabolism, Protein Tyrosine Phosphatases genetics, Protein Tyrosine Phosphatases metabolism
- Abstract
Gene editing of human pluripotent stem cells is a promising approach for developing targeted gene therapies for metabolic diseases. Here, we present a protocol for generating a CRISPR-Cas12a gene knockout of protein tyrosine phosphatases in human embryonic stem cells. We describe steps for differentiating the edited clones into pancreatic islet-like spheroids rich in β-like cells. We then detail procedures for implanting these spheroids under the murine kidney capsule for in vivo maturation., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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