title: Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients.
abstract: Single murine and human intestinal stem cells can be expanded in culture over long time periods as genetically and phenotypically stable epithelial organoids. Increased cAMP levels induce rapid swelling of such organoids by opening the cystic fibrosis transmembrane conductor receptor (CFTR). This response is lost in organoids derived from cystic fibrosis (CF) patients. Here we use the CRISPR/Cas9 genome editing system to correct the CFTR locus by homologous recombination in cultured intestinal stem cells of CF patients. The corrected allele is expressed and fully functional as measured in clonally expanded organoids. This study provides proof of concept for gene correction by homologous recombination in primary adult stem cells derived from patients with a single-gene hereditary defect.
authors: Hubrecht Institute/KNAW, Uppsalalaan 8, Utrecht 3584 CT, The Netherlands; University Medical Center Utrecht, Uppsalalaan 8, Utrecht 3584 CT, The Netherlands.
authors: Gerald
authors: G
authors: Schwank
authors: Bon-Kyoung
authors: BK
authors: Koo
authors: Valentina
authors: V
authors: Sasselli
authors: Johanna F
authors: JF
authors: Dekkers
authors: Inha
authors: I
authors: Heo
authors: Turan
authors: T
authors: Demircan
authors: Nobuo
authors: N
authors: Sasaki
authors: Sander
authors: S
authors: Boymans
authors: Edwin
authors: E
authors: Cuppen
authors: Cornelis K
authors: CK
authors: van der Ent
authors: Edward E S
authors: EE
authors: Nieuwenhuis
authors: Jeffrey M
authors: JM
authors: Beekman
authors: Hans
authors: H
authors: Clevers
mesh_terms: D000328:Adult; D053687:Adult Stem Cells / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D064130:CRISPR-Associated Proteins / Q000378:metabolism*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D015316:Genetic Therapy; D006801:Humans; D007422:Intestines / Q000473:pathology*; D051379:Mice; D008969:Molecular Sequence Data; D009940:Organoids / Q000378:metabolism*; D017393:RNA Editing / Q000235:genetics; D013234:Stem Cells / Q000378:metabolism*
chemical_list: D064130:CRISPR-Associated Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator