TALENs Facilitate Single-step Seamless SDF Correction of F508del CFTR in Airway Epithelial Submucosal Gland Cell-derived CF-iPSCs

Shingo Suzuki; R Geoffrey Sargent; Beate Illek; Horst Fischer; Alaleh Esmaeili-Shandiz; Michael J Yezzi; Albert Lee; Yanu Yang; Soya Kim; Peter Renz; Zhongxia Qi; Jingwei Yu; Marcus O Muench; Ashley I Beyer; Alessander O Guimarães; Lin Ye; Judy Chang; Eli J Fine; Thomas J Cradick; Gang Bao; Meghdad Rahdar; Matthew H Porteus; Tsuyoshi Shuto; Hirofumi Kai; Yuet W Kan; Dieter C Gruenert

doi:10.1038/mtna.2015.43

Molecular Therapy: Nucleic Acids (Jan 2016)

TALENs Facilitate Single-step Seamless SDF Correction of F508del CFTR in Airway Epithelial Submucosal Gland Cell-derived CF-iPSCs

Shingo Suzuki,
R Geoffrey Sargent,
Beate Illek,
Horst Fischer,
Alaleh Esmaeili-Shandiz,
Michael J Yezzi,
Albert Lee,
Yanu Yang,
Soya Kim,
Peter Renz,
Zhongxia Qi,
Jingwei Yu,
Marcus O Muench,
Ashley I Beyer,
Alessander O Guimarães,
Lin Ye,
Judy Chang,
Eli J Fine,
Thomas J Cradick,
Gang Bao,
Meghdad Rahdar,
Matthew H Porteus,
Tsuyoshi Shuto,
Hirofumi Kai,
Yuet W Kan,
Dieter C Gruenert

Affiliations

Shingo Suzuki: Department of Otolaryngology – Head and Neck Surgery, University of California–San Francisco, San Francisco, California, USA
R Geoffrey Sargent: Department of Otolaryngology – Head and Neck Surgery, University of California–San Francisco, San Francisco, California, USA
Beate Illek: Childrens Hospital Oakland Research Institute, Oakland, California, USA
Horst Fischer: Childrens Hospital Oakland Research Institute, Oakland, California, USA
Alaleh Esmaeili-Shandiz: Department of Otolaryngology – Head and Neck Surgery, University of California–San Francisco, San Francisco, California, USA
Michael J Yezzi: Department of Otolaryngology – Head and Neck Surgery, University of California–San Francisco, San Francisco, California, USA
Albert Lee: Department of Otolaryngology – Head and Neck Surgery, University of California–San Francisco, San Francisco, California, USA
Yanu Yang: California Pacific Medical Center Research Institute, San Francisco, California, USA
Soya Kim: Liver Center, University of California–San Francisco, San Francisco, California, USA
Peter Renz: Department of Otolaryngology – Head and Neck Surgery, University of California–San Francisco, San Francisco, California, USA
Zhongxia Qi: Department of Laboratory Medicine, University of California–San Francisco, San Francisco, California, USA
Jingwei Yu: Department of Laboratory Medicine, University of California–San Francisco, San Francisco, California, USA
Marcus O Muench: Department of Laboratory Medicine, University of California–San Francisco, San Francisco, California, USA
Ashley I Beyer: Blood Systems Research Institute, San Francisco, California, USA
Alessander O Guimarães: Blood Systems Research Institute, San Francisco, California, USA
Lin Ye: Department of Medicine, University of California–San Francisco, San Francisco, California, USA
Judy Chang: Department of Medicine, University of California–San Francisco, San Francisco, California, USA
Eli J Fine: Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA
Thomas J Cradick: Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA
Gang Bao: Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA
Meghdad Rahdar: Department of Pediatrics, Stanford University, Stanford, California, USA
Matthew H Porteus: Department of Pediatrics, Stanford University, Stanford, California, USA
Tsuyoshi Shuto: Department of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan
Hirofumi Kai: Department of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan
Yuet W Kan: Department of Medicine, University of California–San Francisco, San Francisco, California, USA
Dieter C Gruenert: Department of Otolaryngology – Head and Neck Surgery, University of California–San Francisco, San Francisco, California, USA

DOI: https://doi.org/10.1038/mtna.2015.43
Journal volume & issue: Vol. 5, no. C

Abstract

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Cystic fibrosis (CF) is a recessive inherited disease associated with multiorgan damage that compromises epithelial and inflammatory cell function. Induced pluripotent stem cells (iPSCs) have significantly advanced the potential of developing a personalized cell-based therapy for diseases like CF by generating patient-specific stem cells that can be differentiated into cells that repair tissues damaged by disease pathology. The F508del mutation in airway epithelial cell-derived CF-iPSCs was corrected with small/short DNA fragments (SDFs) and sequence-specific TALENs. An allele-specific PCR, cyclic enrichment strategy gave ≃100-fold enrichment of the corrected CF-iPSCs after six enrichment cycles that facilitated isolation of corrected clones. The seamless SDF-based gene modification strategy used to correct the CF-iPSCs resulted in pluripotent cells that, when differentiated into endoderm/airway-like epithelial cells showed wild-type (wt) airway epithelial cell cAMP-dependent Cl ion transport or showed the appropriate cell-type characteristics when differentiated along mesoderm/hematopoietic inflammatory cell lineage pathways.

Published in Molecular Therapy: Nucleic Acids

ISSN: 2162-2531 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Medicine: Therapeutics. Pharmacology
Website: https://www.cell.com/molecular-therapy-family/nucleic-acids/latest-content

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