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Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA

Author

Listed:
  • Raphael Haslecker

    (Harvard University)

  • Vincent V. Pham

    (Harvard University)

  • David Glänzer

    (University of Innsbruck)

  • Christoph Kreutz

    (University of Innsbruck)

  • Theodore Kwaku Dayie

    (University of Maryland)

  • Victoria M. D’Souza

    (Harvard University)

Abstract

RNA performs a wide range of functions regulated by its structure, dynamics, and often post-transcriptional modifications. While NMR is the leading method for understanding RNA structure and dynamics, it is currently limited by the inability to reduce spectral crowding by efficient segmental labeling. Furthermore, because of the challenging nature of RNA chemistry, the tools being developed to introduce site-specific modifications are increasingly complex and laborious. Here we use a previously designed Tgo DNA polymerase mutant to present SegModTeX — a versatile, one-pot, copy-and-paste approach to address these challenges. By precise, stepwise construction of a diverse set of RNA molecules, we demonstrate the technique to be superior to RNA polymerase driven and ligation methods owing to its substantially high yield, fidelity, and selectivity. We also show the technique to be useful for incorporating some fluorescent- and a wide range of other probes, which significantly extends the toolbox of RNA biology in general.

Suggested Citation

  • Raphael Haslecker & Vincent V. Pham & David Glänzer & Christoph Kreutz & Theodore Kwaku Dayie & Victoria M. D’Souza, 2023. "Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA," Nature Communications, Nature, vol. 14(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-44254-3
    DOI: 10.1038/s41467-023-44254-3
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    References listed on IDEAS

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    1. Yu Liu & Erik Holmstrom & Jinwei Zhang & Ping Yu & Jinbu Wang & Marzena A. Dyba & De Chen & Jinfa Ying & Stephen Lockett & David J. Nesbitt & Adrian R. Ferré-D’Amaré & Rui Sousa & Jason R. Stagno & Yu, 2015. "Synthesis and applications of RNAs with position-selective labelling and mosaic composition," Nature, Nature, vol. 522(7556), pages 368-372, June.
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