title: TALENs Facilitate Single-step Seamless SDF Correction of F508del CFTR in Airway Epithelial Submucosal Gland Cell-derived CF-iPSCs.
abstract: 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.
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, University of California-San Francisco, San Francisco, California, USA.|Department of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Suzuki
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, University of California-San Francisco, San Francisco, California, USA.|California Pacific Medical Center Research Institute, San Francisco, California, USA.
authors.forename: R Geoffrey
authors.initials: RG
authors.lastname: Sargent
authors.affiliation: Childrens Hospital Oakland Research Institute, Oakland, California, USA.
authors.forename: Beate
authors.initials: B
authors.lastname: Illek
authors.affiliation: Childrens Hospital Oakland Research Institute, Oakland, California, USA.
authors.forename: Horst
authors.initials: H
authors.lastname: Fischer
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, University of California-San Francisco, San Francisco, California, USA.
authors.forename: Alaleh
authors.initials: A
authors.lastname: Esmaeili-Shandiz
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, University of California-San Francisco, San Francisco, California, USA.|California Pacific Medical Center Research Institute, San Francisco, California, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Yezzi
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, University of California-San Francisco, San Francisco, California, USA.|Present address: Graduate Program in Biochemistry, Molecular, Cellular, and Developmental Biology, University of California-Davis, Davis, California, USA.
authors.forename: Albert
authors.initials: A
authors.lastname: Lee
authors.affiliation: California Pacific Medical Center Research Institute, San Francisco, California, USA.|Present address: Molecular Department, Hunter Laboratories, Campbell, California, USA.
authors.forename: Yanu
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Liver Center, University of California-San Francisco, San Francisco, California, USA.|Present address: Heinrich-Heine-Universität Düsseldorf, Institut für Genetik, Düsseldorf, Germany.
authors.forename: Soya
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, University of California-San Francisco, San Francisco, California, USA.|California Pacific Medical Center Research Institute, San Francisco, California, USA.|Present address: Graduate Program in the Department of Biosystems Science and Engineering, ETH, Zürich, Switzerland.
authors.forename: Peter
authors.initials: P
authors.lastname: Renz
authors.affiliation: Department of Laboratory Medicine, University of California-San Francisco, San Francisco, California, USA.
authors.forename: Zhongxia
authors.initials: Z
authors.lastname: Qi
authors.affiliation: Department of Laboratory Medicine, University of California-San Francisco, San Francisco, California, USA.
authors.forename: Jingwei
authors.initials: J
authors.lastname: Yu
authors.affiliation: Department of Laboratory Medicine, University of California-San Francisco, San Francisco, California, USA.|Liver Center, University of California-San Francisco, San Francisco, California, USA.|Blood Systems Research Institute, San Francisco, California, USA.
authors.forename: Marcus O
authors.initials: MO
authors.lastname: Muench
authors.affiliation: Blood Systems Research Institute, San Francisco, California, USA.
authors.forename: Ashley I
authors.initials: AI
authors.lastname: Beyer
authors.affiliation: Blood Systems Research Institute, San Francisco, California, USA.
authors.forename: Alessander O
authors.initials: AO
authors.lastname: Guimarães
authors.affiliation: Department of Medicine, University of California-San Francisco, San Francisco, California, USA.
authors.forename: Lin
authors.initials: L
authors.lastname: Ye
authors.affiliation: Department of Medicine, University of California-San Francisco, San Francisco, California, USA.
authors.forename: Judy
authors.initials: J
authors.lastname: Chang
authors.affiliation: Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
authors.forename: Eli J
authors.initials: EJ
authors.lastname: Fine
authors.affiliation: Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Cradick
authors.affiliation: Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
authors.forename: Gang
authors.initials: G
authors.lastname: Bao
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Meghdad
authors.initials: M
authors.lastname: Rahdar
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Matthew H
authors.initials: MH
authors.lastname: Porteus
authors.affiliation: Department of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Shuto
authors.affiliation: Department of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: Hirofumi
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authors.lastname: Kai
authors.affiliation: Department of Medicine, University of California-San Francisco, San Francisco, California, USA.|Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Helen Diller Family Comprehensive Cancer Center, Institute for Human Genetics, Cardiovascular Research Institute, University of California-San Francisco, San Francisco, California, USA.
authors.forename: Yuet W
authors.initials: YW
authors.lastname: Kan
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, University of California-San Francisco, San Francisco, California, USA.|California Pacific Medical Center Research Institute, San Francisco, California, USA.|Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Helen Diller Family Comprehensive Cancer Center, Institute for Human Genetics, Cardiovascular Research Institute, University of California-San Francisco, San Francisco, California, USA.|Department of Pediatrics, University of Vermont College of Medicine, Burlington, Vermont, USA.
authors.forename: Dieter C
authors.initials: DC
authors.lastname: Gruenert