Selection in vitro of an RNA enzyme that specifically cleaves single-stranded DNA (original) (raw)

Nature volume 344, pages 467–468 (1990)Cite this article

Abstract

THE discovery of RNA enzymes1,2 has, for the first time, provided a single molecule that has both genetic and catalytic properties. We have devised techniques for the mutation, selection and amplification of catalytic RNA, all of which can be performed rapidly _in vitro_3. Here we describe how these techniques can be integrated and performed repeatedly within a single reaction vessel. This allows evolution experiments to be carried out in response to artificially imposed selection constraints. We worked with the Tetrahymena ribozyme, a self-splicing group I intron derived from the large ribosomal RNA precursor of Tetrahymena thermophila that catalyses sequence-specific phosphoester transfer reactions involving RNA substrates4,5. It consists of 413 nucleotides, and assumes a well-defined secondary and tertiary structure responsible for its catalytic activity. We selected for variant forms of the enzyme that could best react with a DNA substrate. This led to the recovery of a mutant form of the enzyme that cleaves DNA more efficiently than the wild-type enzyme. The selected molecule represents the discovery of the first RNA enzyme known to cleave single-stranded DNA specifically.

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Authors and Affiliations

  1. Department of Chemistry and Department of Molecular Biology, Research Institute of Scripps Clinic, 10666 North Torrey Pines Road, La Jolla, California, 92037, USA
    Debra L. Robertson & Gerald F. Joyce

Authors

  1. Debra L. Robertson
  2. Gerald F. Joyce

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Robertson, D., Joyce, G. Selection in vitro of an RNA enzyme that specifically cleaves single-stranded DNA.Nature 344, 467–468 (1990). https://doi.org/10.1038/344467a0

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