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Nat Commun. 2015 Apr 27;6:6952. doi: 10.1038/ncomms7952.

Nanoparticles that deliver triplex-forming peptide nucleic acid molecules correct F508del CFTR in airway epithelium.

Author information

1
Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06520, USA.
2
1] Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06520, USA [2] Yale College, Department of Biomedical Engineering, New Haven, Connecticut 06520, USA.
3
Department of Pediatrics, Yale School of Medicine, New Haven, Connecticut 06510, USA.
4
Department of Cellular and Molecular Physiology, Yale School of Medicine, New Haven, Connecticut 06510, USA.
5
Department of Therapeutic Radiology and Genetics, Yale School of Medicine, New Haven, Connecticut 06510, USA.
6
1] Yale Department of Molecular Biophysics and Biochemistry, New Haven, Connecticut 06520, USA [2] Yale University, Department of Bioinformatics, W.M Keck Foundation Biotechnology Resource Laboratory, New Haven, Connecticut 06511, USA.
7
1] Department of Cellular and Molecular Physiology, Yale School of Medicine, New Haven, Connecticut 06510, USA [2] Department of Surgery, Yale School of Medicine, New Haven, Connecticut 06510, USA.
8
1] Department of Pediatrics, Yale School of Medicine, New Haven, Connecticut 06510, USA [2] Department of Cellular and Molecular Physiology, Yale School of Medicine, New Haven, Connecticut 06510, USA.

Abstract

Cystic fibrosis (CF) is a lethal genetic disorder most commonly caused by the F508del mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. It is not readily amenable to gene therapy because of its systemic nature and challenges including in vivo gene delivery and transient gene expression. Here we use triplex-forming peptide nucleic acids and donor DNA in biodegradable polymer nanoparticles to correct F508del. We confirm modification with sequencing and a functional chloride efflux assay. In vitro correction of chloride efflux occurs in up to 25% of human cells. Deep-sequencing reveals negligible off-target effects in partially homologous sites. Intranasal delivery of nanoparticles in CF mice produces changes in the nasal epithelium potential difference assay, consistent with corrected CFTR function. Also, gene correction is detected in the nasal and lung tissue. This work represents facile genome engineering in vivo with oligonucleotides using a nanoparticle system to achieve clinically relevant levels of gene editing without off-target effects.

PMID:
25914116
PMCID:
PMC4480796
DOI:
10.1038/ncomms7952
[Indexed for MEDLINE]
Free PMC Article

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