Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells

Ana M. Crane, Philipp Kramer, Jacquelin H. Bui, Wook Joon Chung, Xuan Shirley Li, Manuel L. Gonzalez-Garay, Finn Hawkins, Wei Liao, Daniela Mora, Sangbum Choi, Jianbin Wang, Helena C. Sun, David E. Paschon, Dmitry Y. Guschin, Philip D. Gregory, Darrell N. Kotton, Michael C. Holmes, Eric J. Sorscher, Brian R. Davis

Research output: Contribution to journalArticlepeer-review

142 Citations (Scopus)

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 patientderived 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.

Original languageEnglish
Pages (from-to)569-577
Number of pages9
JournalStem Cell Reports
Volume4
Issue number4
DOIs
Publication statusPublished - 2015
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Developmental Biology
  • Cell Biology

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