FACS-assisted CRISPR-Cas9 genome editing of human induced pluripotent stem cells

Titre alternatif
Producteur
Contributeur(s)
Éditeur(s)
Identifiant documentaire 25-4758126
Identifiant OAI 4758126
Auteur(s): Caillaud Amandine,Lévêque Antoine,Thédrez Aurélie,Girardeau Aurore,Canac Robin,Bray Lise,Baudic Manon,Barc Julien,Gaborit Nathalie,Lamirault Guillaume,Gardie Betty,Idriss Salam,Rimbert Antoine,Le May Cédric,Cariou Bertrand,Si-Tayeb Karim
Mots clés CRISPR Flow cytometry Mass cytometry Stem cells
Date de publication 01/12/2022
Date de création
Date de modification
Date d'acceptation du document
Date de dépôt légal
Langue
Thème
Type de ressource
Source
Droits de réutilisation

Région

Département

Commune

Description
This manuscript proposes an efficient and reproducible protocol for the generation of genetically modified human induced pluripotent stem cells (hiPSCs) by genome editing using CRISPR-Cas9 technology. Here, we describe the experimental strategy for generating knockout (KO) and knockin (KI) clonal populations of hiPSCs using single-cell sorting by flow cytometry. We efficiently achieved up to 15 kb deletions, molecular tag insertions, and single-nucleotide editing in hiPSCs. We emphasize the efficacy of this approach in terms of cell culture time. For complete details on the use and execution of this protocol, please refer to Canac et al. (2022) and Bray et al. (2022).
Accès aux documents

0

Consultations

0

Téléchargements