PHOSPHOROTHIOLATE RIBOZYME AGAINST THE CONSERVED SEQUENCE IN V3 LOOP OF HIV-1

Fluorescent-labeled phosphorothiolate DNA-RNA chimeric hammerhead ribo zyme (thio-Rz) was synthesized. The ribozyme recognizes relatively con served sequences in the functionally important region (V3 loop) of env elope protein of HIV-1. The thio-Rz as well as wild-type ribozyme clea ved RNA transcripts (414 base) of V3 region in a dose-dependent manner . After delivery into the cells, both wild-type and thio-Rz were able to be readily detected. The positive signals faded within 12 h in the former case, while they lasted 24 h in the latter case. The inhibitory effect by thio-Rz on cell fusion between envelope-expressing CHO cell s and CD4(+) cells appeared to be time-dependent, and about 70% of inh ibition was seen at 72 h. Possible applications of the ribozyme agains t HIV-1 infection were discussed. (C) 1996 Academic Press, Inc.