Simultaneous high-efficiency base editing and reprogramming of patient fibroblasts

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Jalil , S , Keskinen , T , Maldonado Sartori , R , Sokka , J J , Trokovic , R , Otonkoski , T & Wartiovaara , K 2021 , ' Simultaneous high-efficiency base editing and reprogramming of patient fibroblasts ' , Stem cell reports , vol. 16 , no. 12 , pp. 3064-3075 .

Title: Simultaneous high-efficiency base editing and reprogramming of patient fibroblasts
Author: Jalil, Sami; Keskinen, Timo; Maldonado Sartori, Rocio; Sokka, Juho Joonas; Trokovic, Ras; Otonkoski, Timo; Wartiovaara, Kirmo
Contributor organization: STEMM - Stem Cells and Metabolism Research Program
HUS Children and Adolescents
Date: 2021-11-24
Language: eng
Number of pages: 12
Belongs to series: Stem cell reports
ISSN: 2213-6711
Abstract: 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.
Subject: 1184 Genetics, developmental biology, physiology
1182 Biochemistry, cell and molecular biology
318 Medical biotechnology
Peer reviewed: Yes
Rights: cc_by
Usage restriction: openAccess
Self-archived version: publishedVersion

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