Hampel, Arnold E.
M.S. (Master of Science)
Department of Biological Sciences
Catalytic RNA; HIV (Viruses)
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.
Nesbitt, Steven M., "Targeting an HIV-1 sequence in vivo by the hairpin ribozyme" (1992). Graduate Research Theses & Dissertations. 5209.
vii, 76 pages
Northern Illinois University
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