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Seamless correction of the sickle cell disease mutation of the HBB gene in human induced pluripotent stem cells using TALENs

Authors

  • Ning Sun,

    1. Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois
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  • Huimin Zhao

    Corresponding author
    1. Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois
    2. Departments of Chemical and Biomolecular Engineering, Bioengineering, Chemistry, Center for Biophysics and Computational Biology, Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois
    • Correspondence to: H. Zhao

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  • Conflict of interest statement: None declared.

ABSTRACT

Sickle cell disease (SCD) is the most common human genetic disease which is caused by a single mutation of human β-globin (HBB) gene. The lack of long-term treatment makes the development of reliable cell and gene therapies highly desirable. Disease-specific patient-derived human induced pluripotent stem cells (hiPSCs) have great potential for developing novel cell and gene therapies. With the disease-causing mutations corrected in situ, patient-derived hiPSCs can restore normal cell functions and serve as a renewable autologous cell source for the treatment of genetic disorders. Here we successfully utilized transcription activator-like effector nucleases (TALENs), a recently emerged novel genome editing tool, to correct the SCD mutation in patient-derived hiPSCs. The TALENs we have engineered are highly specific and generate minimal off-target effects. In combination with piggyBac transposon, TALEN-mediated gene targeting leaves no residual ectopic sequences at the site of correction and the corrected hiPSCs retain full pluripotency and a normal karyotype. Our study demonstrates an important first step of using TALENs for the treatment of genetic diseases such as SCD, which represents a significant advance toward hiPSC-based cell and gene therapies. Biotechnol. Biotechnol. Bioeng. 2014;111: 1048–1053. © 2013 Wiley Periodicals, Inc.

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