CD34+ cells from dental pulp stem cells with a ZFN-mediated and homology-driven repair-mediated locus-specific knock-in of an artificial β-globin gene
| Autor: | K Manotham, S Chattong, O Ruangwattanasuk, W Yindeedej, A Setpakdee |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cell Antigens CD34 beta-Globins Biology 03 medical and health sciences stomatognathic system Genome editing hemic and lymphatic diseases Dental pulp stem cells Gene knockin Genetics medicine Humans Molecular Biology Gene Cells Cultured Dental Pulp Endodeoxyribonucleases HEK 293 cells Zinc Fingers Transfection Molecular biology Adult Stem Cells HEK293 Cells 030104 developmental biology medicine.anatomical_structure Molecular Medicine Stem cell Targeted Gene Repair |
| Zdroj: | Gene Therapy. 24:425-432 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/gt.2017.42 |
| Popis: | In humans, mutations in the β-globin gene (HBB) have two important clinical manifestations: β-thalassemia and sickle cell disease. The progress in genome editing and stem cell research may be relevant to the treatment of β-globin-related diseases. In this work, we employed zinc-finger nuclease (ZFN)-mediated gene integration of synthetic β-globin cDNA into HBB loci, thus correcting almost all β-globin mutations. The integration was achieved in both HEK 293 cells and isolated dental pulp stem cell (DPSCs). We also showed that DPSCs with an artificial gene knock-in were capable of generating stable six-cell clones and were expandable at least 108-fold; therefore, they may serve as a personalized stem cell factory. In addition, transfection with non-integrated pCAG-hOct4 and culturing in a conditioned medium converted the genome-edited DPSCs to CD34+ HSC-like cells. We believe that this approach may be useful for the treatment of β-globin-related diseases, especially the severe form of β-thalassemia. |
| Databáze: | OpenAIRE |
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