Cleavage of cottontail rabbit papillomavirus E7 RNA with an anti-E7 ribozyme.

We have designed and constructed a plasmid (pE7RZ-1) that contains a gene for a hammerhead ribozyme that specifically cleaves cottontail rabbit papillomavirus (CRPV) E7 RNA sequences in vitro. The in vitro produced transcripts of pE7RZ-1 cleave CRPV E7 target sequences in trans at 37 degrees C. Pretreatment of pE7RZ-1 RNA with poly-L-lysine (PLL) increases the stability of the RNA to greater than 2 hours in the presence of a crude papilloma cell extract and the pE7RZ-1 RNA/PLL mixture retains catalytic activity in this system. This is the first report of a papillomavirus specific ribozyme RNA cleavage and leads the way for the design of effective papillomavirus specific ribozyme based therapies.