ArticleRNA (New York, N.Y.)2008
Duality of polynucleotide substrates for Phi29 DNA polymerase: 3'-->5' RNase activity of the enzyme.
Article in RNA (New York, N.Y.), 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 25 citations in OpenAlex.
- Programmable AND-gate strategy to reduce false positives in rolling circle amplification.Biosensors & bioelectronics · 2026Article
- USTAT: Unified Sequential Template Amplification and Transcription-a fully synthetic mRNA manufacturing platform.NPJ vaccines · 2026Article
- Reconstituting transcription-translation-coupled DNA replication within complex in vitro biological systems.Nature communications · 2025Article
- One-Pot Detection of miRNA by Dual Rolling Circle Amplification at Ambient Temperature with High Specificity and Sensitivity.Biosensors · 2025Article
- A one-pot isothermal Cas12-based assay for the sensitive detection of microRNAs.Nature biomedical engineering · 2023Article
- Advanced microRNA-based cancer diagnostics using amplified time-gated FRET.Chemical science · 2018Article
- RNase H-assisted RNA-primed rolling circle amplification for targeted RNA sequence detection.Scientific reports · 2018Article
- Dual functional Phi29 DNA polymerase-triggered exponential rolling circle amplification for sequence-specific detection of target DNA embedded in long-stranded genomic DNA.Scientific reports · 2017Article
- Simple methods for the 3' biotinylation of RNA.RNA (New York, N.Y.) · 2014Article
- Amplified microRNA detection by templated chemistry.Nucleic acids research · 2012Article
- Transcript amplification from single bacterium for transcriptome analysis.Genome research · 2011Article
- Direct detection of RNA in vitro and in situ by target-primed RCA: The impact of E. coli RNase III on the detection efficiency of RNA sequences distanced far from the 3'-end.RNA (New York, N.Y.) · 2010Article
- Novel application of Phi29 DNA polymerase: RNA detection and analysis in vitro and in situ by target RNA-primed RCA.RNA (New York, N.Y.) · 2009Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Phi29 DNA polymerase is a small DNA-dependent DNA polymerase that belongs to eukaryotic B-type DNA polymerases. Despite the small size, the polymerase is a multifunctional proofreading-proficient enzyme. It catalyzes two synthetic reactions (polymerization and deoxynucleotidylation of Phi29 terminal protein) and possesses two degradative activities (pyrophosphorolytic and 3'-->5' DNA exonucleolytic activities). Here we report that Phi29 DNA polymerase exonucleolyticaly degrades ssRNA. The RNase activity acts in a 3' to 5' polarity. Alanine replacements in conserved exonucleolytic site (D12A/D66A) inactivated RNase activity of the enzyme, suggesting that a single active site is responsible for cleavage of both substrates: DNA and RNA. However, the efficiency of RNA hydrolysis is approximately 10-fold lower than for DNA. Phi29 DNA polymerase is widely used in rolling circle amplification (RCA) experiments. We demonstrate that exoribonuclease activity of the enzyme can be used for the target RNA conversion into a primer for RCA, thus expanding application potential of this multifunctional enzyme and opening new opportunities for RNA detection.
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