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Use of site directed mutagenesis to analyze function of an anti-HIV Tat ribozyme.

Acquired immune deficiency syndrome (AIDS) is caused by a retrovirus called the Human Immunodeficiency Virus (HIV-1). By 1992, AIDS was the leading cause of death in the US among men aged 22-44 and by January 2000 approximately sixteen million people worldwide had died from this disease. It is estimated 35 million people are living with HIV/AIDS. Current therapies are dedicated to suppressing or inhibiting viral infection of its primary target cell: CD4+ T lymphocytes. These drug therapies do not destroy previously infected cells and are therefore are not curative. It may be possible to use gene therapy to modify target cells and render them resistant to HIV infection. We are currently studying the use of ribozymes as anti-HIV agents. Ribozymes are small catalytic RNAs that specifically cleave single stranded RNA targets. We have designed a hammerhead ribozyme that targets HIV-1 Tat mRNA. Tat is a small virally encoded protein that is responsible for regulation of HIV-1 transcription. Furthermore, Tat interacts directly with its target sequence within the HIV-1 LTR and is therefore critical for viral replication. An anti-Tat ribozyme was synthesized and cloned into both prokaryotic and eukaryotic expression vectors. Initial in vitro testing will be followed by ribozyme analysis in tissue culture models.
Cindy Hollingshead, Dr. William Jackson
Department of Biology-Geology
University of South Carolina, Aiken
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Author:Hollingshead, Cindy; Jackson, William
Publication:Bulletin of the South Carolina Academy of Science
Article Type:Brief Article
Geographic Code:1USA
Date:Jan 1, 2002
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