Poliovirus transcripts with authentic 5' ends generated by a hammerhead ribozyme replicate substantially more rapidly in cell culture without a lag phase compared to transcripts with extra 5' nucleotides.
Poliovirus infectious RNA can be synthesized in vitro using phage DNA-dependent RNA-polymerases. These synthetic transcripts contain several extra nucleotides at the 5' end, which are deleted during replication to generate authentic viral genomes. We removed those 5'-end extra nucleotides utilizing a hammerhead ribozyme to produce transcripts with accurate 5' ends. These transcripts replicate substantially more rapidly in cell culture, demonstrating no lag before replication; they also replicate more efficiently in Xenopus laevis oocytes and in in vitro translation-replication cell extracts. In both systems, an exact 5' end is necessary for synthesis of positive-strand RNA but not negative-strand RNA.
Herold et al. (Sat,) conducted a other in Poliovirus replication (in vitro). Poliovirus transcripts with authentic 5' ends (via hammerhead ribozyme) vs. Poliovirus transcripts with extra 5' nucleotides was evaluated on RNA replication kinetics and positive-strand RNA synthesis. Poliovirus transcripts with authentic 5' ends generated by a hammerhead ribozyme replicate substantially more rapidly in cell culture without a lag phase compared to transcripts with extra 5' nucleotides.