ArticleRNA (New York, N.Y.)2001
Primer-independent RNA sequencing with bacteriophage phi6 RNA polymerase and chain terminators.
Article in RNA (New York, N.Y.), 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
What it found
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Who cites it
9 citing papers in PubMed, 24 citations in OpenAlex.
- Utilization of Bacteriophage phi6 for the Production of High-Quality Double-Stranded RNA Molecules.Viruses · 2024Review
- Applications of phage-derived RNA-based technologies in synthetic biology.Synthetic and systems biotechnology · 2020Review
- Cystoviral RNA-directed RNA polymerases: Regulation of RNA synthesis on multiple time and length scales.Virus research · 2017Review
- RIG-I goes beyond naked recognition.Cell host & microbe · 2013Article
- Insights into the pre-initiation events of bacteriophage phi 6 RNA-dependent RNA polymerase: towards the assembly of a productive binary complex.Nucleic acids research · 2009Article
- Bluetongue virus: dissection of the polymerase complex.The Journal of general virology · 2008Review
- Purified recombinant bluetongue virus VP1 exhibits RNA replicase activity.Journal of virology · 2004Article
- Differential localization of nuclear-encoded tRNAs between the cytosol and mitochondrion in Leishmania tarentolae.RNA (New York, N.Y.) · 2002Article
- Comparison of polymerase subunits from double-stranded RNA bacteriophages.Journal of virology · 2001Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Here we propose a new general method for directly determining RNA sequence based on the use of the RNA-dependent RNA polymerase from bacteriophage phi6 and the chain terminators (RdRP sequencing). The following properties of the polymerase render it appropriate for this application: (1) the phi6 polymerase can replicate a number of single-stranded RNA templates in vitro. (2) In contrast to the primer-dependent DNA polymerases utilized in the sequencing procedure by Sanger et al. (Proc Natl Acad Sci USA, 1977, 74:5463-5467), it initiates nascent strand synthesis without a primer, starting the polymerization on the very 3'-terminus of the template. (3) The polymerase can incorporate chain-terminating nucleotide analogs into the nascent RNA chain to produce a set of base-specific termination products. Consequently, 3' proximal or even complete sequence of many target RNA molecules can be rapidly deduced without prior sequence information. The new technique proved useful for sequencing several synthetic ssRNA templates. Furthermore, using genomic segments of the bluetongue virus we show that RdRP sequencing can also be applied to naturally occurring dsRNA templates. This suggests possible uses of the method in the RNA virus research and diagnostics.
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Registered trials
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