ArticleProceedings of the National Academy of Sciences of the United States of America2005
Design and synthesis of a 3'-O-allyl photocleavable fluorescent nucleotide as a reversible terminator for DNA sequencing by synthesis.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 89 citations in OpenAlex.
- Enzymatic synthesis of RNA oligonucleotides.Nature biotechnology · 2025Article
- Template-independent enzymatic synthesis of RNA oligonucleotides.Nature biotechnology · 2025Article
- The Evolution of Next-Generation Sequencing Technologies.Methods in molecular biology (Clifton, N.J.) · 2025Review
- Controlled enzymatic synthesis of oligonucleotides.Communications chemistry · 2024Review
- Article
- Towards the controlled enzymatic synthesis of LNA containing oligonucleotides.Frontiers in chemistry · 2023Article
- Large-ScaleFrontiers in bioengineering and biotechnology · 2021Review
- Nucleotide analogues as inhibitors of SARS-CoV Polymerase.Pharmacology research & perspectives · 2020Article
- Long walk to genomics: History and current approaches to genome sequencing and assembly.Computational and structural biotechnology journal · 2020Review
- De novo DNA synthesis using polymerase-nucleotide conjugates.Nature biotechnology · 2018Article
- DNA sequencing at 40: past, present and future.Nature · 2017Review
- Multicolor and erasable DNA photolithography.ACS nano · 2014Article
- DNA sequencing by synthesis using 3'-RSC advances · 2014Article
- DNA polymerases drive DNA sequencing-by-synthesis technologies: both past and present.Frontiers in microbiology · 2014Review
- DNA branch migration reactions through photocontrollable toehold formation.Journal of the American Chemical Society · 2013Article
- Photocleavable peptide-oligonucleotide conjugates for protein kinase assays by MALDI-TOF MS.Molecular bioSystems · 2012Article
- Applications of targeted gene capture and next-generation sequencing technologies in studies of human deafness and other genetic disabilities.Hearing research · 2012Review
- Improved nucleotide selectivity and termination of 3'-OH unblocked reversible terminators by molecular tuning of 2-nitrobenzyl alkylated HOMedU triphosphates.Nucleic acids research · 2011Article
- Structural basis for the synthesis of nucleobase modified DNA by Thermus aquaticus DNA polymerase.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Labeled nucleoside triphosphates with reversibly terminating aminoalkoxyl groups.Nucleosides, nucleotides & nucleic acids · 2010Article
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
DNA sequencing by synthesis (SBS) offers an approach for potential high-throughput sequencing applications. In this method, the ability of an incoming nucleotide to act as a reversible terminator for a DNA polymerase reaction is an important requirement to unambiguously determine the identity of the incorporated nucleotide before the next nucleotide is added. A free 3'-OH group on the terminal nucleotide of the primer is necessary for the DNA polymerase to incorporate an incoming nucleotide. Therefore, if the 3'-OH group of an incoming nucleotide is capped by a chemical moiety, it will cause the polymerase reaction to terminate after the nucleotide is incorporated into the DNA strand. If the capping group is subsequently removed to generate a free 3'-OH, the polymerase reaction will reinitialize. We report here the design and synthesis of a 3'-modified photocleavable fluorescent nucleotide, 3'-O-allyl-dUTP-PC-Bodipy-FL-510 (PC-Bodipy, photocleavable 4,4-difluoro-4-bora-3alpha,4alpha-diaza-s-indacene), as a reversible terminator for SBS. This nucleotide analogue contains an allyl moiety capping the 3'-OH group and a fluorophore Bodipy-FL-510 linked to the 5 position of the uracil through a photocleavable 2-nitrobenzyl linker. Here, we have shown that this nucleotide is a good substrate for a DNA polymerase. After the nucleotide was successfully incorporated into a growing DNA strand and the fluorophore was photocleaved, the allyl group was removed by using a Pd-catalyzed reaction to reinitiate the polymerase reaction, thereby establishing the feasibility of using such nucleotide analogues as reversible terminators for SBS.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.