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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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