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      Dynamic and scalable DNA-based information storage

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          Abstract

          The physical architectures of information storage systems often dictate how information is encoded, databases are organized, and files are accessed. Here we show that a simple architecture comprised of a T7 promoter and a single-stranded overhang domain (ss-dsDNA), can unlock dynamic DNA-based information storage with powerful capabilities and advantages. The overhang provides a physical address for accessing specific DNA strands as well as implementing a range of in-storage file operations. It increases theoretical storage densities and capacities by expanding the encodable sequence space and simplifies the computational burden in designing sets of orthogonal file addresses. Meanwhile, the T7 promoter enables repeatable information access by transcribing information from DNA without destroying it. Furthermore, saturation mutagenesis around the T7 promoter and systematic analyses of environmental conditions reveal design criteria that can be used to optimize information access. This simple but powerful ss-dsDNA architecture lays the foundation for information storage with versatile capabilities.

          Abstract

          The physical architectures of information storage dictate how data is encoded, organised and accessed. Here the authors use DNA with a single-strand overhang as a physical address to access specific data and do in-storage file operations in a scalable and reusuable manner.

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          Most cited references32

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          OligoCalc: an online oligonucleotide properties calculator

          We developed OligoCalc as a web-accessible, client-based computational engine for reporting DNA and RNA single-stranded and double-stranded properties, including molecular weight, solution concentration, melting temperature, estimated absorbance coefficients, inter-molecular self-complementarity estimation and intra-molecular hairpin loop formation. OligoCalc has a familiar ‘calculator’ look and feel, making it readily understandable and usable. OligoCalc incorporates three common methods for calculating oligonucleotide-melting temperatures, including a nearest-neighbor thermodynamic model for melting temperature. Since it first came online in 1997, there have been more than 900 000 accesses of OligoCalc from nearly 200 000 distinct hosts, excluding search engines. OligoCalc is available at http://basic.northwestern.edu/biotools/OligoCalc.html, with links to the full source code, usage patterns and statistics at that link as well.
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            Improved thermodynamic parameters and helix initiation factor to predict stability of DNA duplexes.

            To improve the previous DNA/DNA nearest-neighbor parameters, thermodynamic parameters (deltaH degrees, deltaS degrees and deltaG degrees) of 50 DNA/DNA duplexes were measured. Enthalpy change of a helix initiation factor is also considered though the parameters reported recently did not contain the factor. A helix initiation factor for DNA/DNA duplex determined here was the same as that of RNA/RNA duplex (deltaG degrees(37) = 3.4 kcal/mol). The improved nearest-neighbor parameters reproduced not only these 50 experimental values used here but also 15 other experimental values obtained in different studies. Comparing deltaG degrees(37) values of DNA/DNA nearest-neighbor parameters obtained here with those of RNA/RNA and RNA/DNA, RNA/RNA duplex was generally the most stable of the three kinds of duplexes with the same nearest-neighbor sequences. Which is more stable between DNA/DNA and RNA/DNA duplexes is sequence dependent.
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              Initial transcription by RNA polymerase proceeds through a DNA-scrunching mechanism.

              Using fluorescence resonance energy transfer to monitor distances within single molecules of abortively initiating transcription initiation complexes, we show that initial transcription proceeds through a "scrunching" mechanism, in which RNA polymerase (RNAP) remains fixed on promoter DNA and pulls downstream DNA into itself and past its active center. We show further that putative alternative mechanisms for RNAP active-center translocation in initial transcription, involving "transient excursions" of RNAP relative to DNA or "inchworming" of RNAP relative to DNA, do not occur. The results support a model in which a stressed intermediate, with DNA-unwinding stress and DNA-compaction stress, is formed during initial transcription, and in which accumulated stress is used to drive breakage of interactions between RNAP and promoter DNA and between RNAP and initiation factors during promoter escape.
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                Author and article information

                Contributors
                jtuck@ncsu.edu
                ajkeung@ncsu.edu
                Journal
                Nat Commun
                Nat Commun
                Nature Communications
                Nature Publishing Group UK (London )
                2041-1723
                12 June 2020
                12 June 2020
                2020
                : 11
                : 2981
                Affiliations
                [1 ]ISNI 0000 0001 2173 6074, GRID grid.40803.3f, Department of Chemical and Biomolecular Engineering, , North Carolina State University, ; Campus Box 7905, Raleigh, NC 27695-7905 USA
                [2 ]ISNI 0000 0001 2173 6074, GRID grid.40803.3f, Department of Electrical and Computer Engineering, , North Carolina State University, ; Campus Box 7911, Raleigh, NC 27695-7911 USA
                Author information
                http://orcid.org/0000-0001-8975-0294
                http://orcid.org/0000-0001-8958-1232
                Article
                16797
                10.1038/s41467-020-16797-2
                7293219
                32532979
                4af856f5-216b-471a-bb6d-a5c90f5a48bb
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 17 August 2019
                : 20 May 2020
                Funding
                Funded by: FundRef https://doi.org/10.13039/100000144, NSF | Directorate for Computer & Information Science & Engineering | Division of Computer and Network Systems (CNS);
                Award ID: 1650148
                Award ID: 1901324
                Award Recipient :
                Funded by: NSF | Directorate for Computer & Information Science & Engineering | Division of Computer and Network Systems (CNS)
                Funded by: FundRef https://doi.org/10.13039/100007703, North Carolina State University (NC State University);
                Award ID: RISF 2018-2509
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/100005562, North Carolina Biotechnology Center (NCBiotech);
                Award ID: Flash Grant
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/100000138, U.S. Department of Education (ED);
                Award ID: GAANN Fellowship
                Award Recipient :
                Categories
                Article
                Custom metadata
                © The Author(s) 2020

                Uncategorized
                biotechnology,synthetic biology,dna computing and cryptography,chemical engineering,mathematics and computing

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