Sammanfattning
Human induced pluripotent stem cells (hiPSCs) allow in vitro study of genetic diseases and hold potential for personalized stem cell therapy. Gene editing, precisely modifying specifically targeted loci, represents a valuable tool for different hiPSC applications. This is especially useful in monogenic diseases to dissect the function of unknown mutations or to create genetically corrected, patient-derived hiPSCs. Here we describe a highly efficient method for simultaneous base editing and reprogramming of fibroblasts employing a CRISPR-Cas9 adenine base editor. As a proof of concept, we apply this approach to generate gene-edited hiPSCs from skin biopsies of four patients carrying a Finnish-founder pathogenic point mutation in either NOTCH3 or LDLR genes. We also show LDLR activity restoration after the gene correction. Overall, this method yields tens of gene-edited hiPSC monoclonal lines with unprecedented efficiency and robustness while considerably reducing the cell culture time and thus the risk for in vitro alterations.
Originalspråk | engelska |
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Tidskrift | Stem cell reports |
Volym | 16 |
Nummer | 12 |
Sidor (från-till) | 3064-3075 |
Antal sidor | 12 |
ISSN | 2213-6711 |
DOI | |
Status | Publicerad - 24 nov. 2021 |
MoE-publikationstyp | A1 Tidskriftsartikel-refererad |
Vetenskapsgrenar
- 1184 Genetik, utvecklingsbiologi, fysiologi
- 1182 Biokemi, cell- och molekylärbiologi
- 318 Medicinsk bioteknologi