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

Summary

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). Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.

Authors Caillaud A, Lévêque A, Thédrez A, Girardeau A, Canac R, Bray L, Baudic M, Barc J, Gaborit N, Lamirault G, Gardie B, Idriss S, Rimbert A, Le May C, Cariou B, Si-Tayeb K
Journal STAR protocols
Publication Date 2022 Sep 16;3(4):101680
PubMed 36115027
PubMed Central PMC9490201
DOI 10.1016/j.xpro.2022.101680

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