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Efficient and precise CRISPR/Cas9-mediated MECP2 modifications in human-induced pluripotent stem cells

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Item Type:Article
Title:Efficient and precise CRISPR/Cas9-mediated MECP2 modifications in human-induced pluripotent stem cells
Creators Name:Le, T.T.H. and Tran, N.T. and Dao, T.M.L. and Nguyen, D.D. and Do, H.D. and Ha, T.L. and Kühn, R. and Nguyen, T.L. and Rajewsky, K. and Chu, V.T.
Abstract:Patients with Rett syndrome (RTT) have severe mental and physical disabilities. The majority of RTT patients carry a heterozygous mutation in methyl-CpG binding protein 2 (MECP2), an X-linked gene encoding an epigenetic factor crucial for normal nerve cell function. No curative therapy for RTT syndrome exists, and cellular mechanisms are incompletely understood. Here, we developed a CRISPR/Cas9-mediated system that targets and corrects the disease relevant regions of the MECP2 exon 4 coding sequence. We achieved homologous recombination (HR) efficiencies of 20% to 30% in human cell lines and iPSCs. Furthermore, we successfully introduced a MECP2(R270X) mutation into the MECP2 gene in human induced pluripotent stem cells (iPSCs). Consequently, using CRISPR/Cas9, we were able to repair such mutations with high efficiency in human mutant iPSCs. In summary, we provide a new strategy for MECP2 gene targeting that can be potentially translated into gene therapy or for iPSCs-based disease modeling of RTT syndrome.
Keywords:MECP2 Mutations, CRISPR/Cas9, RETT Syndrome, Homologous Recombination, iPSCs
Source:Frontiers in Genetics
ISSN:1664-8021
Publisher:Frontiers Research Foundation (Switzerland)
Volume:10
Page Range:625
Date:July 2019
Official Publication:https://doi.org/10.3389/fgene.2019.00625
PubMed:View item in PubMed

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