Journal article
Partial correction of endogenous ΔF508 CFTR in human cystic fibrosis airway epithelia by spliceosome-mediated RNA trans -splicing
Nature biotechnology, Vol.20(1), pp.47-52
01/2002
DOI: 10.1038/nbt0102-47
PMID: 11753361
Abstract
Spliceosome-mediated RNA trans-splicing (SMaRT) was investigated as a means for functionally correcting endogenous DeltaF508 cystic fibrosis transmembrane conductance regulator (CFTR) transcripts using in vitro human cystic fibrosis (CF) polarized airway epithelia and in vivo human CF bronchial xenografts. Recombinant adenovirus (Ad.CFTR-PTM) encoding a pre-therapeutic molecule (PTM) targeted to CFTR intron 9 corrected transepithelial cyclic AMP (cAMP)-sensitive short-circuit current (Isc) in DeltaF508 homozygous epithelia to a level 16% of that observed in normal human bronchial epithelia. Molecular analyses using RT-PCR and western blotting confirmed SMaRT-mediated partial correction of endogenous DeltaF508 messenger RNA (mRNA) transcripts and protein. In an in vivo model of DeltaF508 CF airway epithelia, human CF bronchial xenografts infected with Ad.CFTR-PTM also demonstrated partial correction of CFTR-mediated Cl- permeability at a level 22% of that seen in non-CF xenografts. These results provide functional evidence for SMaRT-mediated repair of mutant endogenous CFTR mRNA in intact polarized CF airway epithelial models.
Details
- Title: Subtitle
- Partial correction of endogenous ΔF508 CFTR in human cystic fibrosis airway epithelia by spliceosome-mediated RNA trans -splicing
- Creators
- Jonathan A Cohn - Department of Medicine, Duke University Medical CenterMariano A Garcia-Blanco - Intronn IncQinshi Jiang - Department of Anatomy and Cell Biology, College of Medicine, The University of IowaLloyd G Mitchell - Intronn IncXiaoming Liu - Department of Anatomy and Cell Biology, College of Medicine, The University of IowaYulong Zhang - Department of Anatomy and Cell Biology, College of Medicine, The University of Iowa The Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, College of Medicine, The University of IowaS. Gary Mansfield - Intronn IncWeihong Zhou - Department of Anatomy and Cell Biology, College of Medicine, The University of IowaM Puttaraju - Intronn IncJohn F Engelhardt - Department of Anatomy and Cell Biology, College of Medicine, The University of Iowa Department of Internal Medicine, College of Medicine, The University of Iowa The Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, College of Medicine, The University of Iowa
- Resource Type
- Journal article
- Publication Details
- Nature biotechnology, Vol.20(1), pp.47-52
- DOI
- 10.1038/nbt0102-47
- PMID
- 11753361
- ISSN
- 1087-0156
- eISSN
- 1546-1696
- Language
- English
- Date published
- 01/2002
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
- Record Identifier
- 9984025311402771
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