title: Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells.
abstract: Recently developed reprogramming and genome editing technologies make possible the derivation of corrected patient-specific pluripotent stem cell sources-potentially useful for the development of new therapeutic approaches. Starting with skin fibroblasts from patients diagnosed with cystic fibrosis, we derived and characterized induced pluripotent stem cell (iPSC) lines. We then utilized zinc-finger nucleases (ZFNs), designed to target the endogenous CFTR gene, to mediate correction of the inherited genetic mutation in these patient-derived lines via homology-directed repair (HDR). We observed an exquisitely sensitive, homology-dependent preference for targeting one CFTR allele versus the other. The corrected cystic fibrosis iPSCs, when induced to differentiate in vitro, expressed the corrected CFTR gene; importantly, CFTR correction resulted in restored expression of the mature CFTR glycoprotein and restoration of CFTR chloride channel function in iPSC-derived epithelial cells.
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Crane
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Philipp
authors.initials: P
authors.lastname: Kramer
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Jacquelin H
authors.initials: JH
authors.lastname: Bui
authors.affiliation: Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama, Birmingham, AL 35294, USA.
authors.forename: Wook Joon
authors.initials: WJ
authors.lastname: Chung
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Xuan Shirley
authors.initials: XS
authors.lastname: Li
authors.affiliation: Center for Molecular Imaging, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Manuel L
authors.initials: ML
authors.lastname: Gonzalez-Garay
authors.affiliation: Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA 02118, USA.
authors.forename: Finn
authors.initials: F
authors.lastname: Hawkins
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Liao
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Daniela
authors.initials: D
authors.lastname: Mora
authors.affiliation: Division of Clinical and Translational Sciences, Department of Internal Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Sangbum
authors.initials: S
authors.lastname: Choi
authors.affiliation: Sangamo BioSciences, Inc., Richmond, CA 94804, USA.
authors.forename: Jianbin
authors.initials: J
authors.lastname: Wang
authors.affiliation: Sangamo BioSciences, Inc., Richmond, CA 94804, USA.
authors.forename: Helena C
authors.initials: HC
authors.lastname: Sun
authors.affiliation: Sangamo BioSciences, Inc., Richmond, CA 94804, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Paschon
authors.affiliation: Sangamo BioSciences, Inc., Richmond, CA 94804, USA.
authors.forename: Dmitry Y
authors.initials: DY
authors.lastname: Guschin
authors.affiliation: Sangamo BioSciences, Inc., Richmond, CA 94804, USA.
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.affiliation: Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA 02118, USA.
authors.forename: Darrell N
authors.initials: DN
authors.lastname: Kotton
authors.affiliation: Sangamo BioSciences, Inc., Richmond, CA 94804, USA.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.affiliation: Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama, Birmingham, AL 35294, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Sorscher
authors.affiliation: Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA. Electronic address: brian.r.davis@uth.tmc.edu.
authors.forename: Brian R
authors.initials: BR
authors.lastname: Davis
mesh_terms: D000483:Alleles; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000235:genetics* / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics* / Q000378:metabolism*; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018390:Gene Targeting / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D005838:Genotype; D059765:Homologous Recombination; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D009154:Mutation; D059767:Recombinational DNA Repair; D017422:Sequence Analysis, DNA; D016335:Zinc Fingers / Q000235:genetics
chemical_list: D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004720:Endonucleases