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

研究成果: Contribution to journalArticle

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抜粋

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.

元の言語英語
ページ(範囲)225-233
ページ数9
ジャーナルInternational journal of hematology
111
発行部数2
DOI
出版物ステータス出版済み - 2 1 2020

All Science Journal Classification (ASJC) codes

  • Hematology

フィンガープリント CRISPR/Cas9-mediated gene correction in hemophilia B patient-derived iPSCs' の研究トピックを掘り下げます。これらはともに一意のフィンガープリントを構成します。

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    Morishige, S., Mizuno, S., Ozawa, H., Nakamura, T., Mazahery, A., Nomura, K., Seki, R., Mouri, F., Osaki, K., Yamamura, K., Okamura, T., & Nagafuji, K. (2020). CRISPR/Cas9-mediated gene correction in hemophilia B patient-derived iPSCs. International journal of hematology, 111(2), 225-233. https://doi.org/10.1007/s12185-019-02765-0