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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board Member: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Sies, Helmut / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred

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1437-4315
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Volume 386, Issue 2 (Feb 2005)

Issues

Interfering with hepatitis C virus IRES activity using RNA molecules identified by a novel in vitro selection method

Cristina Romero-López
  • Instituto de Parasitología y Biomedicina ‘López-Neyra’, CSIC, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento s/n, Armilla, E-18100 Granada, Spain
/ Alicia Barroso-delJesus
  • Instituto de Parasitología y Biomedicina ‘López-Neyra’, CSIC, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento s/n, Armilla, E-18100 Granada, Spain
/ Elena Puerta-Fernández
  • Instituto de Parasitología y Biomedicina ‘López-Neyra’, CSIC, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento s/n, Armilla, E-18100 Granada, Spain
/ Alfredo Berzal-Herranz
  • Instituto de Parasitología y Biomedicina ‘López-Neyra’, CSIC, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento s/n, Armilla, E-18100 Granada, Spain
Published Online: 2005-06-01 | DOI: https://doi.org/10.1515/BC.2005.023

Abstract

Hepatitis C virus (HCV) infection is one of the world's major health problems, and the identification of efficient HCV inhibitors is a major goal. Here we report the isolation of efficient anti-HCV internal ribosome entry site (IRES) RNA molecules identified by a new in vitro selection method. The newly developed procedure consists of two sequential steps that use distinct criteria for selection: selection for binding and selection for cleaving. The selection protocol was applied to a population of more than 1015 variants of an anti-hepatitis C virus ribozyme covalently linked to an aptamer motif. The ribozyme was directed against positions 357 to 369 of the HCV IRES, and the cleavage substrate was a 691-nucleotide-long RNA fragment that comprises the entire HCV IRES domain. After six selection cycles, seven groups of RNA variants were identified. A representative of each group was tested for its capacity to inhibit IRES activity using in vitro translation assays. All selected RNAs promoted significant inhibition, some by as much as 95%.

Keywords: anti-hepatitis C virus internal ribosome entry site (HCV IRES) RNAs; catalytic RNAs; hepatitis C virus internal ribosome entry site (HCV IRES); inhibitor RNAs; in vitro selection; RNA aptamers

References

About the article

Corresponding author


Received: October 15, 2004

Accepted: December 9, 2004

Published Online: 2005-06-01

Published in Print: 2005-02-01



Citation Information: Biological Chemistry, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/BC.2005.023. Export Citation

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