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Using nucleolytic toxins as restriction enzymes enables new RNA applications

Abstract

Over the past five decades, DNA restriction enzymes have revolutionized biotechnology. While these enzymes are widely used in DNA research and DNA engineering, the emerging field of RNA and mRNA therapeutics requires sequence-specific RNA endoribonucleases. Here, we describe EcoToxN1, a member of the type III toxin-antitoxin family of sequence-specific RNA endoribonucleases, and its use in RNA and mRNA analysis. This enzyme recognizes a specific pentamer in a single-stranded RNA and cleaves the RNA within this sequence. The enzyme is neither dependent on annealing of guide RNA or DNA oligos to the template nor does it require magnesium. Furthermore, it performs over a wide range of temperatures. With its unique functions and characteristics, EcoToxN1 can be classified as an RNA restriction enzyme. EcoToxN1 enables new workflows in RNA analysis and biomanufacturing, meeting the demand for faster, cheaper, and more robust analysis methods.
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Category

Academic article

Language

English

Author(s)

  • Ulli Rothweiler
  • Sigurd Gundesø
  • Emma Wu Mikalsen
  • Steingrim Svenning
  • Mahavir Singh
  • Francis Combes
  • Frida Pettersson
  • Antonia Mangold
  • Yvonne Piotrowski
  • Felix Schwab
  • Olav Lanes
  • Bernd Ketelsen Striberny

Affiliation

  • SINTEF Industry / Biotechnology and Nanomedicine
  • UiT The Arctic University of Norway
  • Indian Institute of Science, Bengaluru
  • ArcticZymes

Year

2024

Published in

Nucleic Acids Research (NAR)

ISSN

0305-1048

Volume

52

Issue

18

View this publication at Norwegian Research Information Repository