ASSURED-optimized CRISPR protocol for knockout/SNP knockin in hiPSCs

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Item Type:	Article
Title:	ASSURED-optimized CRISPR protocol for knockout/SNP knockin in hiPSCs
Creators Name:	Ludwik, K.A., Telugu, N., Schommer, S., Stachelscheid, H. and Diecke, S.
Abstract:	CRISPR-Cas9 technology coupled with human induced pluripotent stem cells allows precise disease modeling in pluripotent cells and subsequently derived specialized cell types. Here, we present an optimized CRISPR-Cas9 pipeline, ASSURED (affordable, successful, specific, user-friendly, rapid, efficient, and deliverable), to produce gene-modified single-cell-derived knockout or single-nucleotide-polymorphism-modified knockin hiPSCs clones. We describe steps for analyzing targeted genomic sequence and designing guide RNAs and homology repair template. We then detail the CRISPR-Cas9 delivery workflow, evaluation of editing efficiency, and automated cell isolation followed by clone screening.
Keywords:	CRISPR-Cas Systems, Gene Editing, Gene Knockout Techniques, Induced Pluripotent Stem Cells, CRISPR-Cas Systems Guide RNA
Source:	STAR Protocols
ISSN:	2666-1667
Publisher:	Elsevier / Cell Press
Volume:	4
Number:	3
Page Range:	102406
Date:	15 September 2023
Official Publication:	https://doi.org/10.1016/j.xpro.2023.102406
PubMed:	View item in PubMed

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