Rotavirus-Like Particles: A Novel Nanocarrier for the Gut

The delivery of bioactive molecules directly to damaged tissues represents a technological challenge. We propose here a new system based on virus-like particles (VLP) from rotavirus, with a marked tropism for the gut to deliver bio-active molecules to intestinal cells. For this, nonreplicative VLP nanoparticles were constructed using a baculovirus expression system and used to deliver an exogenous biomolecule, the green fluorescent protein (GFP), into either MA104 cells or intestinal cells from healthy and 2,4,6-trinitrobenzene sulfonic acid (TNBS)-treated mice. Our results show that expression of rotavirus capsid proteins in baculovirus led to the auto assembly of VLP that display similar properties to rotavirus. In vitro experiments showed that VLP were able to enter into MA104 cells and deliver the reporter protein. Intragastric administration of fluorescent VLP in healthy and TNBS-treated mice resulted in the detection of GFP and viral proteins in intestinal samples. Our results demonstrate an efficient entry of non-replicative rotavirus VLP into the epithelial cell line MA104 and provide the first in vivo evidence of the potential of these nanoparticles as a promising safe candidate for drug delivery to intestinal cells

Identification d'un peptide antiviral contre le virus de l'Hépatite C par une méthode de sélection in cellulo d'éléments génétiques

L'infection par le virus de l'hépatite C (VHC) représente un problème de santé publique. Le VHC est un virus ARN dont la traduction dépend d'un site d'entrée interne du ribosome (IRES) situé dans la région 5' non codante. Afin d'identifier des éléments génétiques du VHC régulateur de sa traduction, nous avons mis en oeuvre une stratégie combinatoire dans un environnement cellulaire. Une banque de fragments aléataoires du génome du VHC a été introduite sous forme de rétrovirus dans des cellules cibles hépatiques exprimant un gène rapporteur sous dépendance de l'IRES du VHC. La sélection des cellules cibles présentant un changement d'expression du rapporteur a permis d'identifier des éléments de la banque potentiellement actifs sur sa traduction. L'étude approfondie d'un élément a conduit à l'identification d'un peptide structuré en hélice a amphiphile, inhibiteur de l'activité de traductionnelle dépendante de l'IRES de VHC et des étapes précoces du cycle viral dans un système de culture cellulaire.The hepatitis C virus (HCV) infection is a major health problem worldwide. HCV is a single stranded, positive sensed RNA virus. Its translation is mediated by an internal ribosomal entry site (IRES) located in the 5' untranslated region. With the aim to identify HCV genetic elements (GE) involved in the control of HCV translation, we developed a combinatorial strategy in cellulo, inspired by the GSE (genetic suppressor elements) technology. A random HCV fragment library was inroduced by retroviral transduction into human hepatic cell lines, expressing a reporter gene under the HCV IRES dependence. The selection of cells with a modified IRES-dependent phenotype led to the identication of several elements presenting some potential activity on HCV translation. We further analyzed the action of one of them and delineated a peptide constituted by the amphipatic a helix, able ro inhibit IRES-dependent translation and HCV infection in a cell culture system.BORDEAUX2-BU Santé (330632101) / SudocSudocFranceF

Sequence Mapping of Combinatorial Libraries on Macro- or Microarrays Experimental Design of DNA Arrays

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