Viral-Mediated Gene Transfer to Study the Molecular Physiology of the Mammalian Inner EarHolt J.R.
Departments of Neuroscience and Otolaryngology, University of Virginia School of Medicine, Charlottesville, Va., USA
Several classes of viral vectors including adenovirus, adeno-associated virus, herpes simplex virus, lentivirus and vaccinia virus have been reported to infect cells of the inner ears of mammals and may be useful for protein manipulation and therapeutic purposes. We have screened a few of these for use as vectors to mediate gene transfer into the sensory hair cells of organotypic cultures from the neonatal mouse cochlea and utricle. Recombinant, replication-deficient adenovirus has emerged as a useful vector for several reasons: ease of vector generation at high titer; efficient hair cell specific infection; robust expression of reporter genes and minimal toxicity. Previously, we characterized adenovirus infected hair cells using a vector that carried the gene for green fluorescent protein (GFP). We screened GFP-positive cells electrophysiologically and found that although hair cells survive adenoviral vector infection, their mechanosensitivity was compromised. Until recently this has limited the scope of adenovirus application to the problems of inner ear physiology and pathophysiology. However, a modified adenoviral vector, now available, has been reported to have reduced ototoxicity in vivo. The modifications include the deletion of the adenoviral genes E1, E3, the viral polymerase, and the preterminal protein. We are currently working to characterize viral-mediated gene transfer into hair cells of the cultured mouse utricle using this new modified adenoviral vector. We have found that hair cells infected with the modified vector have intact hair bundles and robust mechanotransduction.
© 2002 S. Karger AG, Basel
Jeffrey R. Holt, PhD
Assistant Professor, Departments of Neuroscience and Otolaryngology
University of Virginia School of Medicine, Lane Rd.
Charlottesville, VA 22908 (USA)
Tel. +1 434 243 9995, Fax +1 243 982 4380, E-Mail firstname.lastname@example.org
Number of Print Pages : 4
Number of Figures : 3, Number of Tables : 1, Number of References : 14
Audiology & Neuro-Otology (Basic Research and Clinical Applications)
Vol. 7, No. 3, Year 2002 (Cover Date: May-June 2002)
Journal Editor: Manfried Hoke, Münster, Germany
ISSN: 1420–3030 (print), 1421–9700 (Online)
For additional information: http://www.karger.com/journals/aud