Combinatorial selection and edited combinatorial selection of phosphorothioate aptamers targeting human nuclear factor-kappaB RelA/p50 and RelA/RelA.
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.
Bassett
Suzanne E
SE
Department of Human Biological Chemistry and Sealy Center for Structural Biology, Genetics, University of Texas Medical Branch, Galveston, Texas 77555, USA.
Fennewald
Susan M
SM
King
David J
DJ
Li
Xin
X
Herzog
Norbert K
NK
Shope
Robert
R
Aronson
Judy F
JF
Luxon
Bruce A
BA
Gorenstein
David G
DG
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
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