Generation of circular RNAs and trans-cleaving catalytic RNAs by rolling transcription of circular DNA oligonucleotides encoding hairpin ribozymes
Open Access
- 1 July 1998
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 26 (13), 3235-3241
- https://doi.org/10.1093/nar/26.13.3235
Abstract
A simple new strategy for the in vitro synthesis of circular RNAs and hairpin ribozymes is described. Circular single-strand DNA oligonucleotides 67–79 nt in length are constructed to encode both hairpin ribozyme sequences and ribozyme-cleavable sequences. In vitro transcription of these small circles by Escherichia coli RNA polymerase produces long repeating RNAs by a rolling circle mechanism. These repetitive RNAsundergo self-processing, eventually yielding unit length circular and linear RNAs as the chief products. The transcription is efficient despite the absence of promoter sequences, with RNA being produced in up to 400 times the amount of DNA circle used. It is shown that the linear monomeric hairpin ribozymes are active in cleaving RNA targets in trans , including one from HIV-1. Several new findings are established: (i) that rolling circle transcription can be extended to the synthesis of catalytic RNAs outside the hammerhead ribozyme motif; (ii) that rolling circle transcription is potentially a very simple and useful strategy for the generation of circular RNAs in preparative amounts; and (iii) that self-processed hairpin ribozymes can be catalytically active in trans despite the presence of self-binding domains.Keywords
This publication has 41 references indexed in Scilit:
- Peach latent mosaic viroid is locked by a 2′,5′-phosphodiester bond produced by in vitro self-ligationJournal of Molecular Biology, 1997
- Progress Toward the Structure and Therapeutic Use of the Hairpin RibozymeAntisense and Nucleic Acid Drug Development, 1997
- Generation of catalytic RNAs by rolling transcription of synthetic DNA nanocirclesNature Biotechnology, 1997
- Circular Ribozymes Generated in Escherichia coli Using Group I Self-splicing Permuted Intron-Exon SequencesJournal of Biological Chemistry, 1996
- Initiation of Protein Synthesis by the Eukaryotic Translational Apparatus on Circular RNAsScience, 1995
- Antiviral and anticancer ribozymesCurrent Opinion in Oncology, 1994
- The viroid: big punch in a small packageTrends in Microbiology, 1993
- The Single-Nucleotide Addition Cycle in Transcription: a Biophysical and Biochemical PerspectiveAnnual Review of Biophysics and Biophysical Chemistry, 1992
- Sequences required for self-catalysed cleavage of the satellite RNA of tobacco ringspot virusGene, 1989
- RNA catalytic properties of the minimum (-)sTRSV sequenceBiochemistry, 1989