RNA and DNA aptamers in cytomics analysis

RNA and DNA aptamers in cytomics analysis

Autor Ulrich, H. Google Scholar
Martins, Antonio Henrique Baccin Autor UNIFESP Google Scholar
Pesquero, Joao Bosco Autor UNIFESP Google Scholar
Instituição Universidade de São Paulo (USP)
Universidade Federal de São Paulo (UNIFESP)
Resumo Background: the systematic evolution of ligands by exponential enrichment (SELEX) technique is a combinatorial library approach in which DNA or RNA molecules (aptamers) are selected by their ability to bind their protein targets with high affinity and specificity, comparable to that of monoclonal antibodies. in contrast to antibodies conventionally selected in animals, aptamers are generated by an in vitro selection process, and can be directed against almost every target, including antigens like toxins or nonimmunogenic targets, against which conventional antibodies cannot be raised.Methods: Aptamers are ideal candidates for cytomics, as they can be attached to fluorescent reporters or nanoparticles in order to study biological function by fluorescence microscopy, by flow cytometry, or to quantify the concentration of their target in biological fluids or cells using ELISA, RIA, and Western blot assays.Results: We demonstrate the in vitro selection of antikinin B1 receptor aptamers that could be used to determine B1 receptor expression during inflammation processes. These aptamers specifically recognize their target in a Northern-Western blot assay, and bind to their target protein whenever they are exposed in the membrane.Conclusions: Currently, aptamers are linked to fluorescent reporters. We discuss here the present status and future directions concerning the use of the SELEX technique in cytomics. (C) 2004 Wiley-Liss, Inc.
Palavra-chave SELEX
cytomics
fluorescent-tagged aptamers
kinin B1 receptor
Northern-Western-Blot assay
Idioma Inglês
Data de publicação 2004-06-01
Publicado em Cytometry Part A. Hoboken: Wiley-liss, v. 59A, n. 2, p. 220-231, 2004.
ISSN 0196-4763 (Sherpa/Romeo, fator de impacto)
Publicador Wiley-Blackwell
Extensão 220-231
Fonte http://dx.doi.org/10.1002/cyto.a.20056
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000221855700007
Endereço permanente http://repositorio.unifesp.br/handle/11600/27766

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