In vitro characterization of two hairpin ribozymes that cleave HIV-1 RNA
Christ, Wayne A.
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The hairpin ribozyme has been shown to cleave several heterologous RNA targets. In this thesis two hairpin ribozymes were shown in vitro to cleave two separate RNA sequences that are expressed by the virus in vivo. In vitro characterization was performed on these ribozymes to provide kinetic data as well as to optimize the target length. One of the HIV-l sequences is found at position #451 to #465. This targeted region is called TAR and is required for regulation of transcription of all the HIV-1 genes. The other sequence is found at position #6009 to #6024. This region codes for two separate genes called TAT and REV. TAT is a protein that associates with the TAR region to increase the rate of transcription of viral genes. REV is also a protein that associates with a region called the REV responsive element. This association is needed for proper splicing of the viral genes to occur.