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Seamless gene correction of β-thalassemia mutations in patient-specific iPSCs using CRISPR/Cas9 and piggyBac

Xie, Fei, Ye, Lin, Chang, Judy C., Beyer, Ashley I., Wang, Jiaming, Muench, Marcus O., Kan, Yuet Wai·2014·Genome Research
DOI 10.1101/gr.173427.114PMC PMC4158758
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Abstract

β-thalassemia, one of the most common genetic diseases worldwide, is caused by mutations in the human hemoglobin beta ( HBB ) gene. Creation of human induced pluripotent stem cells (iPSCs) from β-thalassemia patients could offer an approach to cure this disease. Correction of the disease-causing mutations in iPSCs could restore normal function and provide a rich source of cells for transplantation. In this study, we used the latest gene-editing tool, CRISPR/Cas9 technology, combined with the piggyBac transposon to efficiently correct the HBB mutations in patient-derived iPSCs without leaving any residual footprint. No off-target effects were detected in the corrected iPSCs, and the cells retain full pluripotency and exhibit normal karyotypes. When differentiated into erythroblasts using a monolayer culture, gene-corrected iPSCs restored expression of HBB compared to the parental iPSCs line. Our study provides an effective approach to correct HBB mutations without leaving any genetic footprint in patient-derived iPSCs, thereby demonstrating a critical step toward the future application of stem cell-based gene therapy to monogenic diseases.

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  • The mutation causing β-thalassaemia could be corrected in human induced pluripotent stem cells ex vivo using CRISPR/Cas9.

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Cited by

Sorted by citations

Authors
Xie, F., Ye, L., Chang, J. C., Beyer, A. I., Wang, J., Muench, M. O., & Kan, Y. W.
Venue
Genome Research
Publisher
Cold Spring Harbor Laboratory
Published
2014-08-05
Volume
24
Issue
9
Pages
1526-1533
ISSN
  • 1088-9051
  • 1549-5469
DOI
10.1101/gr.173427.114
PMC
PMC4158758
Cited by
370
References
5
Language
en
Funders
NIH (P01-DK088760)

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