title: RNA Aptamers as Molecular Tools to Study the Functionality of the Hepatitis C Virus CRE Region.
abstract: BACKGROUND
  Hepatitis C virus (HCV) contains a (+) ssRNA genome with highly conserved structural, functional RNA domains, many of them with unknown roles for the consecution of the viral cycle. Such genomic domains are candidate therapeutic targets. This study reports the functional characterization of a set of aptamers targeting the cis-acting replication element (CRE) of the HCV genome, an essential partner for viral replication and also involved in the regulation of protein synthesis.
  
  
  METHODS
  Forty-four aptamers were tested for their ability to interfere with viral RNA synthesis in a subgenomic replicon system. Some of the most efficient inhibitors were further evaluated for their potential to affect the recruitment of the HCV RNA-dependent RNA polymerase (NS5B) and the viral translation in cell culture.
  
  
  RESULTS
  Four aptamers emerged as potent inhibitors of HCV replication by direct interaction with functional RNA domains of the CRE, yielding a decrease in the HCV RNA levels higher than 90%. Concomitantly, one of them also induced a significant increase in viral translation (>50%). The three remaining aptamers efficiently competed with the binding of the NS5B protein to the CRE.
  
  
  CONCLUSIONS
  Present findings confirm the potential of the CRE as an anti-HCV target and support the use of aptamers as molecular tools for investigating the functionality of RNA domains in viral genomes.
authors.affiliation: Instituto de Parasitología y Biomedicina López-Neyra (IPBLN-CSIC), PTS Granada, Av. Conocimiento, 17, 18016 Armilla, Granada, Spain. alba.fernandez@upf.edu.
authors.forename: Alba
authors.initials: A
authors.lastname: Fernández-Sanlés
authors.affiliation: Instituto de Parasitología y Biomedicina López-Neyra (IPBLN-CSIC), PTS Granada, Av. Conocimiento, 17, 18016 Armilla, Granada, Spain. bbh@ipb.csic.es.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Berzal-Herranz
authors.affiliation: Instituto de Parasitología y Biomedicina López-Neyra (IPBLN-CSIC), PTS Granada, Av. Conocimiento, 17, 18016 Armilla, Granada, Spain. rurik.rgm@gmail.com.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: González-Matamala
authors.affiliation: Instituto de Parasitología y Biomedicina López-Neyra (IPBLN-CSIC), PTS Granada, Av. Conocimiento, 17, 18016 Armilla, Granada, Spain. priosm@ipb.csic.es.
authors.forename: Pablo
authors.initials: P
authors.lastname: Ríos-Marco
authors.affiliation: Instituto de Parasitología y Biomedicina López-Neyra (IPBLN-CSIC), PTS Granada, Av. Conocimiento, 17, 18016 Armilla, Granada, Spain. cristina_romero@ipb.csic.es.
authors.forename: Cristina
authors.initials: C
authors.lastname: Romero-López
authors.affiliation: Instituto de Parasitología y Biomedicina López-Neyra (IPBLN-CSIC), PTS Granada, Av. Conocimiento, 17, 18016 Armilla, Granada, Spain. aberzalh@ipb.csic.es.
authors.forename: Alfredo
authors.initials: A
authors.lastname: Berzal-Herranz
mesh_terms: D052157:Aptamers, Nucleotide / Q000235:genetics*; D002460:Cell Line; D016679:Genome, Viral / Q000235:genetics*; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
chemical_list: D052157:Aptamers, Nucleotide; D012324:RNA-Dependent RNA Polymerase
keywords: 5BSL3.2; CRE; HCV genome; RNA aptamers; antiHCV Aptamers; functional RNA domain