title: Combinatorial selection and edited combinatorial selection of phosphorothioate aptamers targeting human nuclear factor-kappaB RelA/p50 and RelA/RelA.
abstract: Nuclear factor-kappaB (NF-kappaB) transcription factors are important in regulating the immune response and play critical roles in the pathogenesis of chronic inflammatory diseases and a variety of human cancers. Agents that target specific NF-kappaB dimers may serve as therapeutic agents for the prevention of pathogenic immune responses. We have selected monothiophosphate-modified aptamers, or "thioaptamers", to the NF-kappaB p50/RelA heterodimer using combinatorial selection techniques. We also utilized a "double sieve" or editing approach for the generation of thioaptamers with enhanced selectivity to the RelA/RelA homodimer. The thioaptamers from these selections and our previous selections on the p50/p50 and RelA/RelA homodimers all had unique sequences and bound tightly to the recombinant NF-kappaB dimers against which they were selected. The selected thioaptamers also appear to maintain their selectivity and specificity among other cellular proteins, because they have the ability to bind NF-kappaB proteins within nuclear extracts from lipopolysaccharide (LPS)-induced macrophages and B cells.
authors.affiliation: Department of Human Biological Chemistry and Sealy Center for Structural Biology, Genetics, University of Texas Medical Branch, Galveston, Texas 77555, USA.
authors.forename: Suzanne E
authors.initials: SE
authors.lastname: Bassett
authors.forename: Susan M
authors.initials: SM
authors.lastname: Fennewald
authors.forename: David J
authors.initials: DJ
authors.lastname: King
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Norbert K
authors.initials: NK
authors.lastname: Herzog
authors.forename: Robert
authors.initials: R
authors.lastname: Shope
authors.forename: Judy F
authors.initials: JF
authors.lastname: Aronson
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Luxon
authors.forename: David G
authors.initials: DG
authors.lastname: Gorenstein
mesh_terms: D001402:B-Lymphocytes / Q000737:chemistry; D001483:Base Sequence; D002467:Cell Nucleus / Q000737:chemistry; D019281:Dimerization; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D008070:Lipopolysaccharides; D008264:Macrophages / Q000737:chemistry; D016328:NF-kappa B / Q000187:drug effects / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit; D009843:Oligoribonucleotides / Q000737:chemistry / Q000494:pharmacology*; D010710:Phosphates; D051996:Transcription Factor RelA; D014157:Transcription Factors
chemical_list: D008070:Lipopolysaccharides; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; C489548:NFKB1 protein, human; D009843:Oligoribonucleotides; D010710:Phosphates; D051996:Transcription Factor RelA; D014157:Transcription Factors; C035638:thiophosphoric acid