A CRISPR/Cas9-Based Toolkit to Test Gain- and Loss-of-Gene Function in Brain Organoids

Anne Le Digarcher; Céline Lemmers; Arnaud Monteil; Charles Hong; Annie Varrault; Tristan Bouschet

doi:10.1007/978-1-0716-2569-9_5

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A CRISPR/Cas9-Based Toolkit to Test Gain- and Loss-of-Gene Function in Brain Organoids

Anne Le Digarcher, Céline Lemmers, Arnaud Monteil, Charles Hong, Annie Varrault et Tristan Bouschet

Translational Research Methods in Neurodevelopmental Disorders, pp.75-92

Neuromethods, Springer US

30/09/2022

DOI: https://doi.org/10.1007/978-1-0716-2569-9_5

Résumé

Brain

Cortex

CRISPR

CRISPR editing

CRISPRa

CRISPRi

Mouse

Organoids

Pluripotent stem cells

We describe a pipeline to test gene function in brain organoids derived from mouse embryonic stem cells genetically modified with CRISPR/Cas9-based tools. First, we detail methods to affect gene expression in embryonic stem cells by CRISPR activation, CRISPR inhibition, or CRISPR/Cas9 editing, which result in transactivation, repression, and the introduction of a premature termination codon in the gene of interest, respectively. Next, we provide a detailed protocol to differentiate these mutant mouse embryonic stem cells into brain organoids and perform their molecular characterization.

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Détails

Titre: A CRISPR/Cas9-Based Toolkit to Test Gain- and Loss-of-Gene Function in Brain Organoids
Créateurs - sans rôle: Anne Le Digarcher
Céline Lemmers
Arnaud Monteil
Charles Hong
Annie Varrault
Tristan Bouschet
Contributeurs - sans rôle: Stéphane Martin
Frédéric Laumonnier
Détails de publication: Translational Research Methods in Neurodevelopmental Disorders, pp.75-92
Publications en série: Neuromethods
Éditeur: Springer US; New York, NY
Identifiants: 99139452809311
Unité académique: Institut de Génomique Fonctionnelle - IGF
Langue: English
Type de ressource: Book chapter