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.

DOI: 10.1038/mtna.2015.43

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@inproceedings{Suzuki2016TALENsFS, title={TALENs Facilitate Single-step Seamless SDF Correction of F508del CFTR in Airway Epithelial Submucosal Gland Cell-derived CF-iPSCs}, author={Shingo Suzuki and R Geoffrey Sargent and Beate Illek and Horst Fischer and Alaleh Esmaeili-Shandiz and Michael J Yezzi and Albert Lee and Yanu Yang and Soya Kim and Peter Renz and Zhongxia Qi and Jingwei Yu and Marcus O Muench and Ashley I Beyer and Alessander O Guimar{\~a}es and Lin Ye and Judy Chang and Eli J Fine and Thomas J Cradick and Gang Bao and Meghdad Rahdar and Matthew H Porteus and Tsuyoshi Shuto and Hirofumi Kai and Yuet W Kan and Dieter C Gruenert}, booktitle={Molecular therapy. Nucleic acids}, year={2016} }