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 journalArticle

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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 Jan 1
Externally publishedYes

Fingerprint

Induced Pluripotent Stem Cells
Stem cells
Cystic Fibrosis
Genes
Pluripotent Stem Cells
Chloride Channels
Zinc Fingers
Glycoproteins
Fibroblasts
Epithelial Cells
Alleles
Technology
Cell Line
Skin
Mutation
Restoration
Zinc
Repair
Therapeutics
Gene Editing

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Developmental Biology
  • Cell Biology

Cite this

Crane, A. M., Kramer, P., Bui, J. H., Chung, W. J., Li, X. S., Gonzalez-Garay, M. L., ... Davis, B. R. (2015). Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells. Stem Cell Reports, 4(4), 569-577. https://doi.org/10.1016/j.stemcr.2015.02.005

Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells. / Crane, Ana M.; Kramer, Philipp; Bui, Jacquelin H.; Chung, Wook Joon; Li, Xuan Shirley; Gonzalez-Garay, Manuel L.; Hawkins, Finn; Liao, Wei; Mora, Daniela; Choi, Sangbum; Wang, Jianbin; Sun, Helena C.; Paschon, David E.; Guschin, Dmitry Y.; Gregory, Philip D.; Kotton, Darrell N.; Holmes, Michael C.; Sorscher, Eric J.; Davis, Brian R.

In: Stem Cell Reports, Vol. 4, No. 4, 01.01.2015, p. 569-577.

Research output: Contribution to journalArticle

Crane, AM, Kramer, P, Bui, JH, Chung, WJ, Li, XS, Gonzalez-Garay, ML, Hawkins, F, Liao, W, Mora, D, Choi, S, Wang, J, Sun, HC, Paschon, DE, Guschin, DY, Gregory, PD, Kotton, DN, Holmes, MC, Sorscher, EJ & Davis, BR 2015, 'Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells', Stem Cell Reports, vol. 4, no. 4, pp. 569-577. https://doi.org/10.1016/j.stemcr.2015.02.005
Crane, Ana M. ; Kramer, Philipp ; Bui, Jacquelin H. ; Chung, Wook Joon ; Li, Xuan Shirley ; Gonzalez-Garay, Manuel L. ; Hawkins, Finn ; Liao, Wei ; Mora, Daniela ; Choi, Sangbum ; Wang, Jianbin ; Sun, Helena C. ; Paschon, David E. ; Guschin, Dmitry Y. ; Gregory, Philip D. ; Kotton, Darrell N. ; Holmes, Michael C. ; Sorscher, Eric J. ; Davis, Brian R. / Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells. In: Stem Cell Reports. 2015 ; Vol. 4, No. 4. pp. 569-577.
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