Construction and selection of bead-bound combinatorial oligonucleoside phosphorothioate and phosphorodithioate aptamer libraries designed for rapid PCR-based sequencing.

Xianbin Yang, Suzanne E. Bassett, Xin Li, Bruce A. Luxon, Norbert K. Herzog, Robert E. Shope, Judy Aronson, Tarl W. Prow, James F. Leary, Romy Kirby, Andrew D. Ellington, David G. Gorenstein

Research output: Contribution to journalArticle

55 Scopus citations

Abstract

Chemically synthesized combinatorial libraries of unmodified or modified nucleic acids have not previously been used in methods to rapidly select oligonucleotides binding to target biomolecules such as proteins. Phosphorothioate oligonucleotides (S-ODNs) or phosphorodithioate oligonucleotides (S2-ODNs) with sulfurs replacing one or both of the non-bridging phosphate oxygens bind to proteins more tightly than unmodified oligonucleotides and have the potential to be used as diagnostic reagents and therapeutics. We have applied a split synthesis methodology to create one-bead one-S-ODN and one-bead one-S2-ODN libraries. Binding and selection of specific beads to the transcription factor NF-kappaB p50/p50 protein were demonstrated. Sequencing both the nucleic acid bases and the positions of any 3'-O-thioate/dithioate linkages was carried out by using a novel PCR-based identification tag of the selected beads. This approach allows us to rapidly and conveniently identify S-ODNs or S2-ODNs that bind to proteins.

Original languageEnglish (US)
Pages (from-to)e132
JournalNucleic acids research
Volume30
Issue number23
DOIs
StatePublished - Dec 1 2002

ASJC Scopus subject areas

  • Genetics

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    Yang, X., Bassett, S. E., Li, X., Luxon, B. A., Herzog, N. K., Shope, R. E., Aronson, J., Prow, T. W., Leary, J. F., Kirby, R., Ellington, A. D., & Gorenstein, D. G. (2002). Construction and selection of bead-bound combinatorial oligonucleoside phosphorothioate and phosphorodithioate aptamer libraries designed for rapid PCR-based sequencing. Nucleic acids research, 30(23), e132. https://doi.org/10.1093/nar/gnf132