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Efficient gene editing of human induced pluripotent stem cells using CRISPR/Cas9

Item Type:Article
Title:Efficient gene editing of human induced pluripotent stem cells using CRISPR/Cas9
Creators Name:Yumlu, S. and Bashir, S. and Stumm, J. and Kühn, R.
Abstract:The generation of targeted mutants is a crucial step toward studying the biomedical effect of genes of interest. The generation of such mutants in human induced pluripotent stem cells (iPSCs) is of an utmost importance as these cells carry the potential to be differentiated into any cell lineage. Using the CRISPR/Cas9 nuclease system for induction of targeted double-strand breaks, gene editing of target loci in iPSCs can be achieved with high efficiency. This chapter covers protocols for the preparation of reagents to target loci of interest, the transfection, and for the genotyping of single cell-derived iPSC clones. Furthermore, we provide a protocol for the convenient generation of plasmids enabling multiplex gene targeting.
Keywords:Pluripotent Stem Cells, Gene Editing, CRISPR, Cas9, Knockout, Knockin, i53, Trex2
Source:Methods in Molecular Biology
Series Name:Methods in Molecular Biology
Title of Book:CRISPR Gene Editing : Methods and Protocols
ISSN:1064-3745
ISBN:978-1-4939-9169-3
Publisher:Springer / Humana Press (U.S.A.)
Volume:1961
Page Range:137-151
Date:2019
Official Publication:https://doi.org/10.1007/978-1-4939-9170-9_10
PubMed:View item in PubMed

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