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Protocol for CRISPR-Cas12a genome editing of protein tyrosine phosphatases in human pluripotent stem cells and functional β-like cell generation

par Negueruela Escudero, Javier ;Vandenbempt, Valerie ;Talamantes, Stephanie ;Ribeiro-Costa, Francisco;Nunes, Mariana;Marques Dias, André ;Bansal, Mayank ;Gurzov, Esteban Nicolas
Référence STAR Protocols, 5, 3, page (103297)
Publication Publié, 2024-09-01

Article révisé par les pairs

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Résumé :

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.