Work

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

Year: 2024

Type: article

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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103297

Authors: Javier Negueruela, Valerie Vandenbempt, Stephanie Talamantes, Francisco Ribeiro-Costa, Mariana Nunes, André Dias, Mayank Bansal, Esteban N. Gurzov

Institution: Université Libre de Bruxelles

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,500

Funders: U.S. Department of Agriculture, Helsingin Yliopisto, Juvenile Diabetes Research Foundation United States of America, Fonds De La Recherche Scientifique - FNRS, European Research Council, H2020 European Research Council, JDRF

Awards: G318251033, G4592458386, PROTEIN TYROSINE PHOSPHATASES IN METABOLIC DISEASES: OXIDATION, DYSFUNCTION AND THERAPEUTIC POTENTIAL

OpenAlex ID: W4402292185

FWCI: 0.579

Cites: 33

Cited by: 5

Related to: 10