Journal article
Transcriptional Targeting in the Airway Using Novel Gene Regulatory Elements
American journal of respiratory cell and molecular biology, Vol.47(2), pp.227-233
08/2012
DOI: 10.1165/rcmb.2011-0444OC
PMCID: PMC3423461
PMID: 22447971
Abstract
The delivery of cystic fibrosis transmembrane conductance regulator (CFTR) to airway epithelia is a goal of many gene therapy strategies to treat cystic fibrosis. Because the native regulatory elements of the
CFTR
are not well characterized, the development of vectors with heterologous promoters of varying strengths and specificity would aid in our selection of optimal reagents for the appropriate expression of the vector-delivered
CFTR
gene. Here we contrasted the performance of several novel gene-regulatory elements. Based on airway expression analysis, we selected putative regulatory elements from
BPIFA1
and
WDR65
to investigate. In addition, we selected a human
CFTR
promoter region (∼ 2 kb upstream of the human
CFTR
transcription start site) to study. Using feline immunodeficiency virus vectors containing the candidate elements driving firefly luciferase, we transduced murine nasal epithelia
in vivo
. Luciferase expression persisted for 30 weeks, which was the duration of the experiment. Furthermore, when the nasal epithelium was ablated using the detergent polidocanol, the mice showed a transient loss of luciferase expression that returned 2 weeks after administration, suggesting that our vectors transduced a progenitor cell population. Importantly, the
hWDR65
element drove sufficient
CFTR
expression to correct the anion transport defect in
CFTR
-null epithelia. These results will guide the development of optimal vectors for sufficient, sustained
CFTR
expression in airway epithelia.
Details
- Title: Subtitle
- Transcriptional Targeting in the Airway Using Novel Gene Regulatory Elements
- Creators
- Erin R Burnight - Interdisciplinary Graduate Program in GeneticsGuoshun Wang - Gene Therapy Program, Department of Microbiology, Department of Genetics, and Department of Medicine, Louisiana State University Health Sciences Center, New Orleans, LouisianaPaul B McCray - Interdisciplinary Graduate Program in GeneticsPatrick L Sinn - Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, and
- Resource Type
- Journal article
- Publication Details
- American journal of respiratory cell and molecular biology, Vol.47(2), pp.227-233
- DOI
- 10.1165/rcmb.2011-0444OC
- PMID
- 22447971
- PMCID
- PMC3423461
- NLM abbreviation
- Am J Respir Cell Mol Biol
- ISSN
- 1044-1549
- eISSN
- 1535-4989
- Publisher
- American Thoracic Society
- Language
- English
- Date published
- 08/2012
- Academic Unit
- Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Internal Medicine; Ophthalmology and Visual Sciences
- Record Identifier
- 9984093337802771
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