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PMID: 9113977 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A general purpose RNA-cleaving DNA enzyme.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 94 ·No. 9 ·1997-04-29 ·Pages 4262-6

Santoro SW, Joyce GF

Abstract

An in vitro selection procedure was used to develop a DNA enzyme that can be made to cleave almost any targeted RNA substrate under simulated physiological conditions. The enzyme is comprised of a catalytic domain of 15 deoxynucleotides, flanked by two substrate-recognition domains of seven to eight deoxynucleotides each. The RNA substrate is bound through Watson-Crick base pairing and is cleaved at a particular phosphodiester located between an unpaired purine and a paired pyrimidine residue. Despite its small size, the DNA enzyme has a catalytic efficiency (kcat/Km) of approximately 10(9) M-1.min-1 under multiple turnover conditions, exceeding that of any other known nucleic acid enzyme. Its activity is dependent on the presence of Mg2+ ion. By changing the sequence of the substrate-recognition domains, the DNA enzyme can be made to target different RNA substrates. In this study, for example, it was directed to cleave synthetic RNAs corresponding to the start codon region of HIV-1 gag/pol, env, vpr, tat, and nef mRNAs.

Keywords
Non-programmatic
MeSH Terms
DNA, Catalytic DNA, Single-Stranded/metabolism Genes, Viral HIV-1 Oligodeoxyribonucleotides/metabolism Oligoribonucleotides/metabolism RNA, Catalytic/metabolism RNA, Messenger/metabolism RNA, Viral/metabolism Selection, Genetic
Chemicals
DNA, Catalytic DNA, Single-Stranded Oligodeoxyribonucleotides Oligoribonucleotides RNA, Catalytic RNA, Messenger RNA, Viral RNA-cleaving DNA 10-23
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Santoro S W
Department of Chemistry, Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Joyce G F
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.94.9.4262
Published
1997-04-29
Pages
4262-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC20710
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.94.9.4262 PMC Full Text View journal details
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