A shortened adeno-associated virus expression cassette for CFTR gene transfer to cystic fibrosis airway epithelia

Lynda S Ostedgaard; Tatiana Rokhlina; Philip H Karp; Philip Lashmit; Sandra Afione; Michael Schmidt; Joseph Zabner; Mark F Stinski; Jay A Chiorini; Michael J Welsh

doi:10.1073/pnas.0409845102

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A shortened adeno-associated virus expression cassette for CFTR gene transfer to cystic fibrosis airway epithelia

Journal article

Open access

Peer reviewed

A shortened adeno-associated virus expression cassette for CFTR gene transfer to cystic fibrosis airway epithelia

Lynda S Ostedgaard, Tatiana Rokhlina, Philip H Karp, Philip Lashmit, Sandra Afione, Michael Schmidt, Joseph Zabner, Mark F Stinski, Jay A Chiorini and Michael J Welsh

Proceedings of the National Academy of Sciences - PNAS, Vol.102(8), pp.2952-2957

02/22/2005

DOI: 10.1073/pnas.0409845102

PMCID: PMC549485

PMID: 15703296

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Abstract

Adeno-associated viruses (AAVs) such as AAV5 that transduce airway epithelia from the apical surface are attractive vectors for gene transfer in cystic fibrosis (CF). However, their utility in CF has been limited because packaging of the insert becomes inefficient when its length exceeds ≈4,900–5,000 bp. To partially circumvent this size constraint, we previously developed a CF transmembrane conductance regulator (CFTR) transgene that deleted a portion of the R domain (CFTRΔR). In this study, we focused on shortening the other elements in the AAV expression cassette. We found that portions of the CMV immediate/early ( CMVie ) enhancer/promoter could be deleted without abolishing activity. We also tested various intervening sequences, poly(A) signals, and an intron to develop an expression cassette that meets the size restrictions imposed by AAV. We then packaged these shortened elements with the CFTRΔR transgene into AAV5 and applied them to the apical surface of differentiated CF airway epithelia. Two to 4 weeks later, the AAV5 vectors partially corrected the CF Cl – transport defect. These results demonstrate that a single AAV vector can complement the CF defect in differentiated airway epithelia and thereby further the development of effective CF gene transfer.

Biological Sciences

gene therapy

promoter

CMV

Details

Title: Subtitle: A shortened adeno-associated virus expression cassette for CFTR gene transfer to cystic fibrosis airway epithelia
Creators: Lynda S Ostedgaard - Howard Hughes Medical Institute and Departments of
Tatiana Rokhlina - Howard Hughes Medical Institute and Departments of
Philip H Karp - Howard Hughes Medical Institute and Departments of
Philip Lashmit - Howard Hughes Medical Institute and Departments of
Sandra Afione - Howard Hughes Medical Institute and Departments of
Michael Schmidt - Howard Hughes Medical Institute and Departments of
Joseph Zabner - Howard Hughes Medical Institute and Departments of
Mark F Stinski - Howard Hughes Medical Institute and Departments of
Jay A Chiorini - Howard Hughes Medical Institute and Departments of
Michael J Welsh - Howard Hughes Medical Institute and Departments of
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.102(8), pp.2952-2957
DOI: 10.1073/pnas.0409845102
PMID: 15703296
PMCID: PMC549485
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences
Language: English
Date published: 02/22/2005
Academic Unit: Neurology; Molecular Physiology and Biophysics; Pulmonary, Critical Care, and Occupational Medicine; Microbiology and Immunology; Center for Biocatalysis and Bioprocessing; Neurosurgery; Internal Medicine
Record Identifier: 9984020614102771

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