CRISPR/Cas9-mediated gene correction in hemophilia B patient-derived iPSCs.
Int J Hematol
; 111(2): 225-233, 2020 Feb.
Article
em En
| MEDLINE
| ID: mdl-31664646
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.
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Base de dados:
MEDLINE
Assunto principal:
Terapia Genética
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Hemofilia B
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Células-Tronco Pluripotentes Induzidas
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Sistemas CRISPR-Cas
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Edição de Genes
Limite:
Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article