Functional Repair of CFTR by CRISPR/Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients

被引：1068

作者：

Schwank, Gerald ^{[1
,2
]}

Koo, Bon-Kyoung ^{[1
,2
]}

Sasselli, Valentina ^{[1
,2
]}

Dekkers, Johanna F. ^{[3
,4
]}

Heo, Inha ^{[1
,2
]}

Demircan, Turan ^{[1
]}

Sasaki, Nobuo ^{[1
,2
]}

Boymans, Sander ^{[1
]}

Cuppen, Edwin ^{[1
,6
]}

van der Ent, Cornelis K. ^{[3
]}

Nieuwenhuis, Edward E. S. ^{[5
]}

Beekman, Jeffrey M. ^{[5
,6
]}

Clevers, Hans ^{[1
,2
]}

机构：

[1] Hubrecht Inst KNAW, NL-3584 CT Utrecht, Netherlands

[2] Univ Med Ctr Utrecht, NL-3584 CT Utrecht, Netherlands

[3] Wilhelmina Childrens Hosp, Univ Med Ctr, Dept Pediat Pulmonol, NL-3584 EA Utrecht, Netherlands

[4] Wilhelmina Childrens Hosp, Univ Med Ctr, Dept Immunol, NL-3584 EA Utrecht, Netherlands

[5] Wilhelmina Childrens Hosp, Univ Med Ctr, Dept Pediat Gastroenterol, NL-3584 EA Utrecht, Netherlands

[6] UMC Utrecht, Dept Med Genet, NL-3584 GG Utrecht, Netherlands

来源：

CELL STEM CELL | 2013年 / 13卷 / 06期

基金：

欧洲研究理事会; 英国惠康基金;

关键词：

ZINC-FINGER NUCLEASES; IN-VITRO EXPANSION; RNA-GUIDED ENDONUCLEASE; HOMOLOGOUS RECOMBINATION; CAS9; NUCLEASE; WNT RECEPTORS; GENE; DNA; IDENTIFICATION; SPECIFICITY;

D O I：

10.1016/j.stem.2013.11.002

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

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.

引用

页码：653 / 658

页数：6

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