CRISPR/Cas9-mediated gene correction in hemophilia B patient-derived iPSCs

Satoshi Morishige, Shinichi Mizuno, Hidetoshi Ozawa, Takayuki Nakamura, Ahmad Mazahery, Kei Nomura, Ritsuko Seki, Fumihiko Mouri, Koichi Osaki, Kenichi Yamamura, Takashi Okamura, Koji Nagafuji

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)


The clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system is an efficient genome-editing tool that holds potential for gene therapy. Here, we report an application of this system for gene repair in hemophilia B (HB) using induced pluripotent stem cells (iPSCs). We prepared targeting plasmids with homology arms containing corrected sequences to repair an in-frame deletion in exon 2 of the factor IX (F9) gene and transfected patient-derived iPSCs with the Cas9 nuclease and a guide RNA expression vector. To validate the expression of corrected F9, we attempted to induce the differentiation of iPSCs toward hepatocyte-like cells (HLCs) in vitro. We successfully repaired a disease-causing mutation in HB in patient-derived iPSCs. The transcription product of corrected F9 was confirmed in HLCs differentiated from gene-corrected iPSCs. Although further research should be undertaken to obtain completely functional hepatocytes with secretion of coagulation factor IX, our study provides a proof-of-principle for HB gene therapy using the CRISPR/Cas9 system.

Original languageEnglish
Pages (from-to)225-233
Number of pages9
JournalInternational journal of hematology
Issue number2
Publication statusPublished - Feb 1 2020

All Science Journal Classification (ASJC) codes

  • Hematology


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