ArticleProceedings of the National Academy of Sciences of the United States of America2016
Design and characterization of a nanopore-coupled polymerase for single-molecule DNA sequencing by synthesis on an electrode array.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
27 citing papers in PubMed.
- G-HIV: An Integrated Long-Read Sequencing and Automated Bioinformatics Platform for Rapid and Precise HIV-1 Surveillance.Microorganisms · 2026Article
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- Thermodynamic Profiling Reveals DNA Polymerase Template Binding, Substrate Incorporation, and Exonuclease Function.International journal of molecular sciences · 2025Article
- Cryogenic light microscopy of vitrified samples with angstrom precision.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
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- Decoding information stored in DNAs through single-molecule electrical detection platforms.Innovation (Cambridge (Mass.)) · 2025Review
- Characterization and Engineering of Two Novel Strand-Displacing B Family DNA Polymerases fromBiomolecules · 2025Article
- MtlA gene sequencing based on 3rd -generation sequencing for Vibrio typing.BMC microbiology · 2025Article
- Expression and functional study of DNA polymerases fromFrontiers in microbiology · 2024Article
- Engineering psychrophilic polymerase for nanopore long-read sequencing.Frontiers in bioengineering and biotechnology · 2024Article
- Unraveling the salt tolerance of Phi29 DNA polymerase using compartmentalized self-replication and microfluidics platform.Frontiers in microbiology · 2023Article
- Molecular electronics sensors on a scalable semiconductor chip: A platform for single-molecule measurement of binding kinetics and enzyme activity.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Detection of single peptide with only one amino acid modification via electronic fingerprinting using reengineered durable channel of Phi29 DNA packaging motor.Biomaterials · 2021Article
- Optimized cDICE for Efficient Reconstitution of Biological Systems in Giant Unilamellar Vesicles.ACS synthetic biology · 2021Article
- Uncertainties in synthetic DNA-based data storage.Nucleic acids research · 2021Review
- Power to the protein: enhancing and combining activities using the Spy toolbox.Chemical science · 2020Review
- Cell Separations and Sorting.Analytical chemistry · 2020Review
- Solid-state nanopores towards single-molecule DNA sequencing.Journal of human genetics · 2020Review
- Nanopore device-based fingerprinting of RNA oligos and microRNAs enhanced with an Osmium tag.Scientific reports · 2019Article
- A Nanopore Approach for Analysis of Caspase-7 Activity in Cell Lysates.Biophysical journal · 2019Article
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Authors and funding
30 authors.
Funding
Abstract
Scalable, high-throughput DNA sequencing is a prerequisite for precision medicine and biomedical research. Recently, we presented a nanopore-based sequencing-by-synthesis (Nanopore-SBS) approach, which used a set of nucleotides with polymer tags that allow discrimination of the nucleotides in a biological nanopore. Here, we designed and covalently coupled a DNA polymerase to an α-hemolysin (αHL) heptamer using the SpyCatcher/SpyTag conjugation approach. These porin-polymerase conjugates were inserted into lipid bilayers on a complementary metal oxide semiconductor (CMOS)-based electrode array for high-throughput electrical recording of DNA synthesis. The designed nanopore construct successfully detected the capture of tagged nucleotides complementary to a DNA base on a provided template. We measured over 200 tagged-nucleotide signals for each of the four bases and developed a classification method to uniquely distinguish them from each other and background signals. The probability of falsely identifying a background event as a true capture event was less than 1.2%. In the presence of all four tagged nucleotides, we observed sequential additions in real time during polymerase-catalyzed DNA synthesis. Single-polymerase coupling to a nanopore, in combination with the Nanopore-SBS approach, can provide the foundation for a low-cost, single-molecule, electronic DNA-sequencing platform.
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Registered trials
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.