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Development of receptor-based inhibitory RNA aptamers for anthrax toxin neutralization.
Lee, Sang-Choon; Gedi, Vinayakumar; Ha, Na-Reum; Cho, Jun-Haeng; Park, Hae-Chul; Yoon, Moon-Young.
Afiliação
  • Lee SC; Department of Chemistry and Research Institute of Natural Sciences, Hanyang University, Seoul 133-791, Republic of Korea.
  • Gedi V; Department of Chemistry and Research Institute of Natural Sciences, Hanyang University, Seoul 133-791, Republic of Korea.
  • Ha NR; Department of Chemistry and Research Institute of Natural Sciences, Hanyang University, Seoul 133-791, Republic of Korea.
  • Cho JH; Department of Chemistry and Research Institute of Natural Sciences, Hanyang University, Seoul 133-791, Republic of Korea.
  • Park HC; Veterinary Drugs & Biologics Division, Animal and Plant Quarantine Agency (QIA), Anyang 430-757, Republic of Korea.
  • Yoon MY; Department of Chemistry and Research Institute of Natural Sciences, Hanyang University, Seoul 133-791, Republic of Korea. Electronic address: myyoon@hanyang.ac.kr.
Int J Biol Macromol ; 77: 293-302, 2015.
Article em En | MEDLINE | ID: mdl-25841381
ABSTRACT
Anthrax toxin excreted by Bacillus anthracis is the key causative agent of infectious anthrax disease. In the present study, we targeted the binding of PA to the ATR/TEM8 Von Willebrand factor type A (VWA) domain, which we cloned into Escherichia coli and purified to homogeneity under denaturing conditions. To develop an anthrax toxin inhibitor, we selected and identified short single strand RNA aptamers (approximately 30mer) consisting of different sequences of nucleic acids with a high binding affinity in the 100 nanomolar range against the recombinant ATR/TEM8 VWA domain using systematic evolution of ligands by exponential enrichment (SELEX). Five candidate aptamers were further characterized by several techniques including secondary structural analysis. The inhibitor efficiency (IC50) of one of the aptamers toward anthrax toxin was approximately 5µM in macrophage RAW 264.7 cells, as determined from cytotoxicity analysis by MTT assay. We believe that the candidate aptamers should be useful for blocking the binding of PA to its receptor in order to neutralize anthrax toxin.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Toxinas Bacterianas / Receptores de Superfície Celular / Aptâmeros de Nucleotídeos / Técnica de Seleção de Aptâmeros / Antígenos de Bactérias / Proteínas de Neoplasias Tipo de estudo: Prognostic_studies Limite: Animals / Humans Idioma: En Revista: Int J Biol Macromol Ano de publicação: 2015 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Toxinas Bacterianas / Receptores de Superfície Celular / Aptâmeros de Nucleotídeos / Técnica de Seleção de Aptâmeros / Antígenos de Bactérias / Proteínas de Neoplasias Tipo de estudo: Prognostic_studies Limite: Animals / Humans Idioma: En Revista: Int J Biol Macromol Ano de publicação: 2015 Tipo de documento: Article