Publication: Correction of Hemoglobin E/Beta-Thalassemia Patient-Derived iPSCs Using CRISPR/Cas9
20
Issued Date
2021-01-01
Resource Type
ISSN
19406029
10643745
10643745
Other identifier(s)
2-s2.0-85097997794
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Methods in Molecular Biology. Vol.2211, (2021), 193-211
Suggested Citation
Methichit Wattanapanitch Correction of Hemoglobin E/Beta-Thalassemia Patient-Derived iPSCs Using CRISPR/Cas9. Methods in Molecular Biology. Vol.2211, (2021), 193-211. doi:10.1007/978-1-0716-0943-9_14 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/76423
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Correction of Hemoglobin E/Beta-Thalassemia Patient-Derived iPSCs Using CRISPR/Cas9
Author(s)
Other Contributor(s)
Abstract
HbE/β-thalassemia is one of the most common thalassemic syndromes in Southeast Asia and Thailand. Patients have mutations in β hemoglobin (HBB) gene resulting in decreased and/or abnormal production of β hemoglobin. Here, we describe a protocol for CRISPR/Cas9-mediated gene correction of the mutated hemoglobin E from one allele of the HBB gene by homology-directed repair (HDR) in HbE/β-thalassemia patient-derived induced pluripotent stem cells (iPSCs) using a CRISPR/Cas9 plasmid-based transfection method and a single-stranded DNA oligonucleotide (ssODN) repair template harboring the correct nucleotides. Our strategy allows the seamless HbE gene correction with the editing efficiency (HDR) up to 3%, as confirmed by Sanger sequencing. This protocol provides a simple one-step genetic correction of HbE mutation in the patient-derived iPSCs. Further differentiation of the corrected iPSCs into hematopoietic stem/progenitor cells will provide an alternative renewable source of cells for the application in autologous transplantation in the future.