Enhancement and inhibition by 2′-O-hydroxyethyl residues of gene targeting mediated by triple helix forming oligonucleotides

Mrinalkanti Kundu, Fumi Nagatsugi, Alokes Majumdar, Paul S. Miller, Michael M. Seidman

Research output: Contribution to journalArticlepeer-review

Abstract

Reagents that recognize and bind specific genomic sequences in living mammalian cells would have great potential for genetic manipulation, including gene knockout, strain construction, and gene therapy. Triple helix forming oligonucleotides (TFOs) bind specific sequences via the major groove, but pyrimidine motif TFOs are limited by their poor activity under physiological conditions. Base and sugar analogues that overcome many of these limitations have been described. In particular, 2′-O-modifications influence sugar pucker and third strand conformation, and have been important to the development of bioactive TFOs. Here we have analyzed the impact of 2′-O-hydroxyethyl (2′-HE) substitutions, in combination with other 2′ modifications. We prepared modified TFOs conjugated to psoralen and measured targeting activity in a gene knockout assay in cultured hamster cells. We find that 2′-HE residues enhance the bioactivity of TFOs containing 2′-O-methyl (2′-OMe) modifications, but reduce the bioactivity of TFOs containing, in addition, 2′-O-aminoethyl (2′-AE) residues.

Original languageEnglish (US)
Pages (from-to)1927-1938
Number of pages12
JournalNucleosides, Nucleotides and Nucleic Acids
Volume22
Issue number10
DOIs
StatePublished - 2003

Keywords

  • Bioactivity
  • Oligonucleotides
  • Ribose analogue
  • Triple helix

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Genetics

Fingerprint

Dive into the research topics of 'Enhancement and inhibition by 2′-O-hydroxyethyl residues of gene targeting mediated by triple helix forming oligonucleotides'. Together they form a unique fingerprint.

Cite this