Generation of nuclease resistant circular RNA decoys for HIV-Tat and HIV-Rev by autocatalytic splicing.
Circular exon sequences can be generated by splicing permuted intron-exon (PIE) sequences. The Anabaena pre-tRNA group I self-splicing PIE sequence was modified to generate circular forms of the HIV-TAR and the high affinity region of the HIV-RRE (RBE). RNA products containing TAR and the RBE were purified from splicing reactions and demonstrated to be circular. The circular form of these sequences was shown to be resistant to nuclease degradation in cellular extracts. Gel shift assays demonstrate that the circular form of the RBE is specifically bound by a Rev derived peptide. These data suggest that PIE-circularization of RNA may be an effective way to express small stable RNAs designed for therapeutics (eg. decoys).