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

• Published in: Biotechnology and bioengineering 2014-05, Vol.111 (5), p.1048-1053
• Main Authors: Sun, Ning, Zhao, Huimin
• Format: Article
• Language: English
• Subjects: Anemia, Sickle Cell - genetics; beta-Globins - genetics; Biotechnology; Cells, Cultured; Cloning, Molecular - methods; Deoxyribonucleases - genetics; Deoxyribonucleases - metabolism; Gene therapy; Genetic disorders; Genetic Therapy; genome editing; Humans; Induced Pluripotent Stem Cells; Mutation; piggyBac transposon; Sickle cell disease; Stem cells; TAL effector nucleases
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.25018

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. By using an engineered TALEN and piggyBac transposon, the authors successfully corrected the sickle cell disease mutation of the HBB gene in patient‐derived induced pluripotent stem cells in a specific and seamless manner. This study demonstrates an important first step of using TALENs for the treatment of genetic diseases, which represents a significant advance toward stem cell and gene therapies.