Novel combinatorial selection of phosphorothioate oligonucleotide aptamers

David J. King, Daniel A. Ventura, Allan R. Brasier, David G. Gorenstein

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

A novel combinatorial approach is described for construction and screening of enhanced nuclease-resistant phosphorothioate DNA 'decoys' or 'aptamers.' Aptamers have been selected that bind tightly to the nuclear factor for human IL6 (NF-IL6), a basic leucine zipper transcription factor involved in the induction of acute-phase responsive and cytokine gene promotors in response to inflammation. Using a random combinatorial selection approach and dNTP(αS)'s in the PCR amplification, we can select specific thio-substituted agents which have the highest specificity in binding to target NF-IL6. Using a 22-nucleotide-long duplex random library, nanomolar binding, specific 22-mer thiophosphate backbone substitution sequences (at dA positions only) were selected. These show a different consensus sequence than normal phosphate backbone CCAAT/enhancer binding protein recognition sites for NF-IL6. Unlike the wild-type 10-mer sequences, which bind 1 protein dimer/duplex, these 22-mer thiophosphate aptamers bind with a stoichiometry of 2 dimers/duplex.

Original languageEnglish (US)
Pages (from-to)16489-16493
Number of pages5
JournalBiochemistry
Volume37
Issue number47
DOIs
StatePublished - Nov 24 1998

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Phosphorothioate Oligonucleotides
Interleukin-6
Dimers
Basic-Leucine Zipper Transcription Factors
CCAAT-Enhancer-Binding Proteins
Consensus Sequence
Stoichiometry
Libraries
Amplification
Screening
Substitution reactions
Nucleotides
Genes
Phosphates
Cytokines
Inflammation
Polymerase Chain Reaction
DNA
Proteins
thiophosphoric acid

ASJC Scopus subject areas

  • Biochemistry

Cite this

King, D. J., Ventura, D. A., Brasier, A. R., & Gorenstein, D. G. (1998). Novel combinatorial selection of phosphorothioate oligonucleotide aptamers. Biochemistry, 37(47), 16489-16493. https://doi.org/10.1021/bi981780f

Novel combinatorial selection of phosphorothioate oligonucleotide aptamers. / King, David J.; Ventura, Daniel A.; Brasier, Allan R.; Gorenstein, David G.

In: Biochemistry, Vol. 37, No. 47, 24.11.1998, p. 16489-16493.

Research output: Contribution to journalArticle

King, DJ, Ventura, DA, Brasier, AR & Gorenstein, DG 1998, 'Novel combinatorial selection of phosphorothioate oligonucleotide aptamers', Biochemistry, vol. 37, no. 47, pp. 16489-16493. https://doi.org/10.1021/bi981780f
King DJ, Ventura DA, Brasier AR, Gorenstein DG. Novel combinatorial selection of phosphorothioate oligonucleotide aptamers. Biochemistry. 1998 Nov 24;37(47):16489-16493. https://doi.org/10.1021/bi981780f
King, David J. ; Ventura, Daniel A. ; Brasier, Allan R. ; Gorenstein, David G. / Novel combinatorial selection of phosphorothioate oligonucleotide aptamers. In: Biochemistry. 1998 ; Vol. 37, No. 47. pp. 16489-16493.
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