Abstract

application): The goal of this pilot project is to explore the potential of RNA replication for prolonging the synthesis of messenger RNAs in the context of gene therapy for cystic fibrosis. RNA replication will be catalyzed by the RNA-dependent RNA polymerase that is encoded by one of the nodaviruses, a family of small, spherical, RNA viruses of insects, with bipartite single-stranded RNA genomes. The larger segment of the viral genome replicates autonomously when introduced into a wide variety of cells, and also supports the replication of other RNAs that carry appropriate cis-acting signals. In recent work, we used a hormone-inducible RNA polymerase II promoter to initiate vigorous RNA replication from the nuclei of cultured cells transfected with the RNA replicase cDNA. We have now designed self-contained DNA cassettes to launch replication of the mRNAs for several different reporter genes. The levels and duration of reporter gene expression will be compared with and without mRNA replication, in cultured cells and in the lungs of mice, thereby allowing us to determine the contribution of RNA replication to the abundance and longevity of gene expression in these two situations. Finally, we will attempt to achieve the replicative amplification of CFTR mRNA in this system and examine its expression in cultured cells, including primary airway cells from CF patients.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Center Core Grants (P30)
Project #
5P30DK054781-04
Application #
6564367
Study Section
Project Start
2002-01-01
Project End
2002-12-31
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
4
Fiscal Year
2002
Total Cost
$165,355
Indirect Cost

  Institution

Name
University of Alabama Birmingham
Department
Type
DUNS #
004514360
City
Birmingham
State
AL
Country
United States
Zip Code
35294

  Publications

Blount, Angela; Zhang, Shaoyan; Chestnut, Michael et al. (2011) Transepithelial ion transport is suppressed in hypoxic sinonasal epithelium. Laryngoscope 121:1929-34
Havasi, Viktoria; Rowe, Steven M; Kolettis, Peter N et al. (2010) Association of cystic fibrosis genetic modifiers with congenital bilateral absence of the vas deferens. Fertil Steril 94:2122-7
Su, Xuefeng; Li, Qingnan; Shrestha, Kedar et al. (2006) Interregulation of proton-gated Na(+) channel 3 and cystic fibrosis transmembrane conductance regulator. J Biol Chem 281:36960-8
Sorscher, E J; Harris, J; Alexander, M et al. (2006) Activators of viral gene expression in polarized epithelial monolayers identified by rapid-throughput drug screening. Gene Ther 13:781-8
Joung, Insil; Harber, Greg; Gerecke, Kimberly M et al. (2005) Improved gene delivery into neuroglial cells using a fiber-modified adenovirus vector. Biochem Biophys Res Commun 328:1182-7
Bebok, Zsuzsa; Collawn, James F; Wakefield, John et al. (2005) Failure of cAMP agonists to activate rescued deltaF508 CFTR in CFBE41o- airway epithelial monolayers. J Physiol 569:601-15
Joung, Insil; Kim, Hak Jae; Kwon, Yunhee Kim (2005) p62 modulates Akt activity via association with PKCzeta in neuronal survival and differentiation. Biochem Biophys Res Commun 334:654-60
Yoshino, Naoto; Lu, Fabien X-S; Fujihashi, Kohtaro et al. (2004) A novel adjuvant for mucosal immunity to HIV-1 gp120 in nonhuman primates. J Immunol 173:6850-7
Hardiman, Karin M; McNicholas-Bevensee, Carmel M; Fortenberry, James et al. (2004) Regulation of amiloride-sensitive Na(+) transport by basal nitric oxide. Am J Respir Cell Mol Biol 30:720-8
Burrus, Jason K; Matheson, William D; Hong, Jeong S et al. (2004) Hepatocyte growth factor stimulates adenoviral-mediated gene transfer across the apical membrane of epithelial cells. J Gene Med 6:624-30

Showing the most recent 10 out of 57 publications