Publication Date

1992

Document Type

Dissertation/Thesis

First Advisor

Hampel, Arnold E.

Degree Name

M.S. (Master of Science)

Department

Department of Biological Sciences

LCSH

Catalytic RNA||HIV (Viruses)

Abstract

Ribozymes are small catalytic RNAs that can be engineered to cleave heterologous RNA in an enzymatic fashion. One of the applications of ribozyme technology is as an AIDS therapeutic. In this thesis, an HIV-1 sequence was targeted by the hairpin ribozyme in vivo. A unique sequence found at position +111/112 of the 5' leader sequence, which is part of the 5' LTR, was targeted. This domain is needed for the transactivation and translation of the HIV-1 genome, without which the virus would not be able to replicate. The chloramphenicol acetyl transferase gene (CAT) served as a reporter and was placed under control of the 5' LTR of HIV-1. Constructs were stably integrated into Hela T4+ cells and CAT activity was monitored before and after induction of the ribozyme, which was under the control of the mouse mammary tumor virus promoter (MMTV). A 41% reduction in CAT activity was observed 12 hours after ribozyme induction. Due to the instability of the Hela T4+ genome, further testing was not possible to verify that the reduction of CAT activity was due to catalysis by the ribozyme or was caused by the ribozyme acting as an antisense molecule.

Comments

Includes bibliographical references (pages [69]-76)

Extent

vii, 76 pages

Language

eng

Publisher

Northern Illinois University

Rights Statement

In Copyright

Rights Statement 2

NIU theses are protected by copyright. They may be viewed from Huskie Commons for any purpose, but reproduction or distribution in any format is prohibited without the written permission of the authors.

Media Type

Text

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