| RRC ID |
58135
|
| 著者 |
Morishige S, Mizuno S, Ozawa H, Nakamura T, Mazahery A, Nomura K, Seki R, Mouri F, Osaki K, Yamamura K, Okamura T, Nagafuji K.
|
| タイトル |
CRISPR/Cas9-mediated gene correction in hemophilia B patient-derived iPSCs.
|
| ジャーナル |
Int J Hematol
|
| Abstract |
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.
|
| 巻・号 |
111(2)
|
| ページ |
225-233
|
| 公開日 |
2020-2-1
|
| DOI |
10.1007/s12185-019-02765-0
|
| PII |
10.1007/s12185-019-02765-0
|
| PMID |
31664646
|
| MeSH |
CRISPR-Cas Systems / genetics*
Gene Editing*
Genetic Therapy / methods*
Hemophilia B / genetics*
Hemophilia B / therapy*
Humans
Induced Pluripotent Stem Cells*
|
| IF |
2.245
|
| 引用数 |
1
|
| リソース情報 |
| ヒト・動物細胞 |
201B7(HPS0063) |
