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CRISPR/Cas9-mediated gene correction in hemophilia B patient-derived iPSCs
- Source :
- International Journal of Hematology. 111:225-233
- Publication Year :
- 2019
- Publisher :
- Springer Science and Business Media LLC, 2019.
-
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.
- Subjects :
- Gene Editing
Expression vector
Cas9
Genetic enhancement
Induced Pluripotent Stem Cells
Genetic Therapy
Hematology
Biology
Hemophilia B
Molecular biology
03 medical and health sciences
Exon
0302 clinical medicine
030220 oncology & carcinogenesis
medicine
Humans
CRISPR
CRISPR-Cas Systems
Induced pluripotent stem cell
Gene
030215 immunology
Factor IX
medicine.drug
Subjects
Details
- ISSN :
- 18653774 and 09255710
- Volume :
- 111
- Database :
- OpenAIRE
- Journal :
- International Journal of Hematology
- Accession number :
- edsair.doi.dedup.....d8acf8eb66cb48e8aa86aa541b3c67e9