Nature Methods
- 4, 733 - 739 (2007)
Published online: 5 August 2007; | doi:10.1038/nmeth1077
An HSV vector system for selection of ligand-gated ion channel modulatorsRahul Srinivasan1, Shaohua Huang1, Suchita Chaudhry1, Adrian Sculptoreanu2, David Krisky3, Michael Cascio1, Peter A Friedman2, William C de Groat2, Darren Wolfe1 & Joseph C Glorioso11
Department of Molecular Genetics and Biochemistry, 200 Lothrop Street, Biomedical Science Tower, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA. 2
Department of Pharmacology, 200 Lothrop Street, Biomedical Science Tower, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA. 3
Department of Pathology, 200 Lothrop Street, Biomedical Science Tower, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
Correspondence should be addressed to Joseph C Glorioso glorioso@pitt.edu Pathological alterations of ion channel activity result from changes in modulatory mechanisms governing receptor biology. Here we describe a conditional herpes simplex virus (HSV) replication-based strategy to discover channel modulators whereby inhibition of agonist-induced channel activation by a vector-expressed modulatory gene product prevents ion flux, osmotic shock and cell death. Inhibition of channel activity, in this case, the rat vanilloid (Trpv1 or the glycine receptor (GlyR 1), can allow selection of escape vector plaques containing the 'captured' modulatory gene for subsequent identification and functional analysis. We validated this prediction using mixed infections of a wild-type Trpv1 expression vector vTTHR and a nonfunctional 'poreless' Trpv1 subunit-expressing vector, vHP, wherein vHP was highly selected from a large background of vTTHR viruses in the presence of the Trpv1 agonist, capsaicin. The approach should be useful for probing large libraries of vector-expressed cDNAs for the presence of ion channel modulators.
MORE ARTICLES LIKE THIS These links to content published by NPG are automatically generated.
|
