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Targeting sites within HIV-1 cDNA with a DNA-cleaving ribozyme

Academic Article
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Overview

authors

  • Raillard, S. A.
  • Joyce, Gerald

publication date

  • 1996

journal

  • Biochemistry  Journal

abstract

  • A variant of the Tetrahymena ribozyme that efficiently cleaves single-stranded DNA under simulated physiological conditions [Tsang, J., & Joyce, G. F. (1994) Biochemistry 33, 5966-5973] was evaluated as a potential therapeutic agent on the basis of its ability to cleave synthetic oligonucleotide substrates corresponding to conserved target sites within HIV-I cDNA. In order to increase the sequence selectivity of the ribozyme, its substrate recognition domain was extended from 6 to 12 nucleotides, allowing base pairing with substrate nucleotides that lie both upstream and downstream of the cleavage site. The sequence of the extended recognition domain could be changed to allow cleavage of a variety of different DNA targets. The ribozyme exhibited a high degree of sequence specificity, discriminating by a factor of 10(2) to more than 10(4) against substrates that form a single-base mismatch with the ribozyme's recognition domain. Mismatches that occurred close to the cleavage site led to a greater decrease in activity compared to those that occurred farther away.

subject areas

  • Animals
  • Base Composition
  • Base Sequence
  • DNA Primers
  • DNA, Complementary
  • DNA, Viral
  • Directed Molecular Evolution
  • HIV-1
  • Kinetics
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Oligodeoxyribonucleotides
  • RNA, Catalytic
  • Repetitive Sequences, Nucleic Acid
  • Substrate Specificity
  • Tetrahymena
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Identity

International Standard Serial Number (ISSN)

  • 0006-2960

Digital Object Identifier (DOI)

  • 10.1021/bi960845g

PubMed ID

  • 8794750
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Additional Document Info

start page

  • 11693

end page

  • 11701

volume

  • 35

issue

  • 36

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