Rolling circle RNA synthesis catalysed by RNA

  1. Emil Laust Kristoffersen
  2. Matthew Burman
  3. Agnes Noy
  4. Philipp Holliger  Is a corresponding author
  1. MRC Laboratory of Molecular Biology, United Kingdom
  2. University of York, United Kingdom

Abstract

RNA-catalysed RNA replication is widely considered a key step in the emergence of life's first genetic system. However, RNA replication can be impeded by the extraordinary stability of duplex RNA products, which must be dissociated for re-initiation of the next replication cycle. Here we have explored rolling circle synthesis (RCS) as a potential solution to this strand separation problem. We observe sustained RCS by a triplet polymerase ribozyme beyond full-length circle synthesis with strand displacement yielding concatemeric RNA products. Furthermore, we show RCS of a circular Hammerhead ribozyme capable of self-cleavage and re-circularisation. Thus, all steps of a viroid-like RNA replication pathway can be catalysed by RNA alone. Finally, we explore potential RCS mechanisms by molecular dynamics simulations, which indicate a progressive build-up of conformational strain upon RCS' with destabilisation of nascent strand 5'- and 3'-ends. Our results have implications for the emergence of RNA replication and for understanding the potential of RNA to support complex genetic processes.

Data availability

All data generated or analyzed in this manuscript is supplied within the manuscript or supporting file; Source Data files containing original unedited gels images as well as numeric data have been provided for Figures 1,2,4 and 5, as well as figure supplements when relevant.Modelling data and sequencing data are provided as described in the data availability section in the manuscript.

The following data sets were generated

Article and author information

Author details

  1. Emil Laust Kristoffersen

    MRC Laboratory of Molecular Biology, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8965-8201
  2. Matthew Burman

    Department of Physics, University of York, York, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Agnes Noy

    Department of Physics, University of York, York, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0673-8949
  4. Philipp Holliger

    MRC Laboratory of Molecular Biology, Cambridge, United Kingdom
    For correspondence
    ph1@mrc-lmb.cam.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3440-9854

Funding

Carlsbergfondet (CF17-0809)

  • Emil Laust Kristoffersen

Medical Research Council (MC_U105178804)

  • Philipp Holliger

Engineering and Physical Sciences Research Council (EP/N027639/1)

  • Agnes Noy

Engineering and Physical Sciences Research Council (EP/R513386/1)

  • Matthew Burman

Engineering and Physical Sciences Research Council (EP/T022205/1)

  • Agnes Noy

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Timothy W Nilsen, Case Western Reserve University, United States

Version history

  1. Received: November 1, 2021
  2. Preprint posted: November 30, 2021 (view preprint)
  3. Accepted: February 1, 2022
  4. Accepted Manuscript published: February 2, 2022 (version 1)
  5. Version of Record published: March 21, 2022 (version 2)
  6. Version of Record updated: March 22, 2022 (version 3)

Copyright

© 2022, Kristoffersen et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Emil Laust Kristoffersen
  2. Matthew Burman
  3. Agnes Noy
  4. Philipp Holliger
(2022)
Rolling circle RNA synthesis catalysed by RNA
eLife 11:e75186.
https://doi.org/10.7554/eLife.75186

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https://doi.org/10.7554/eLife.75186

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