Combinatorial Selection and Edited Combinatorial Selection of Phosphorothioate Aptamers Targeting Human Nuclear Factor-kappaB RelA/p50 and RelA/RelA

摘要

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.