Preparation of HIV TAR RNA with RNA scissors

Two hammerhead ribozymes derived from plant pathogenic RNAs were used to cu t off the HIV TAR RNA from the T7 RNA transcript through a cis cleavage rea ction, Stem I of the (+) vLTSV ribozyme comprises 8 nucleotides of the 5' t erminus of TAR RNA, but stem III of the (+)sTRSV ribozyme consists of 8 nuc leotides of the 3' end of TAR RNA, The construct containing two GUC hammerh ead ribozyme target sequences identified the cleavage sites to cut off a re quired RNA molecule. This method was applied for preparation of 35 nt long TAR, RNA. Its activity was proved by the complex formation with, the Tat pr otein. It seems that this approach based on RNA scissors can also be used f or the generation of required RNA molecules, RNA decoys or RNA aptamers in vivo.