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


  • T.T.H. Le
  • N.T. Tran
  • T.M.L. Dao
  • D.D. Nguyen
  • H.D. Do
  • T.L. Ha
  • R. Kühn
  • T.L. Nguyen
  • K. Rajewsky
  • V.T. Chu


  • Frontiers in Genetics


  • Front Genet 10: 625


  • 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.