Identification of a methylated oligoribonucleotide as a potent inhibitor of HIV-1 reverse transcription complex

Upon HIV-1 infection of a target cell, the viral reverse transcriptase (RT) copies the genomic RNA to synthesize the viral DNA. The genomic RNA is within the incoming HIV-1 core where it is coated by molecules of nucleocapsid (NC) protein that chaperones the reverse transcription process. Indeed, the RT chaperoning properties of NC extend from the initiation of cDNA synthesis to completion of the viral DNA. New and effective drugs against HIV-1 continue to be required, which prompted us to search for compounds aimed at inhibiting NC protein. Here, we report that the NC chaperoning activity is extensively inhibited in vitro by small methylated oligoribonucleotides (mODN). These mODNs were delivered intracellularly using a cell-penetrating-peptide and found to impede HIV-1 replication in primary human cells at nanomolar concentrations. Extensive analysis showed that viral cDNA synthesis was severely impaired by mODNs. Partially resistant viruses with mutations in NC and RT emerged after months of passaging in cell culture. A HIV-1 molecular clone (NL4.3) bearing these mutations was found to replicate at high concentrations of mODN, albeit with a reduced fitness. Small, methylated ODNs such as mODN-11 appear to be a new type of highly potent inhibitor of HIV-1

Management de la qualité dans les laboratoires d'analyses de biologie médicale (proposition d'un programme d'évaluation)

AIX-MARSEILLE2-BU Pharmacie (130552105) / SudocSudocFranceF

Transfert de gènes dans le modèle murin de la maladie de Sandhoff à l'aide de vecteurs lentiviraux

PARIS7-Bibliothèque centrale (751132105) / SudocSudocFranceF

Imaging Minimal Bacteria at the Nanoscale: a Reliable and Versatile Process to Perform Single-Molecule Localization Microscopy in Mycoplasmas

Mycoplasmas are the smallest free-living organisms. These bacteria are important models for both fundamental and synthetic biology, owing to their highly reduced genomes. They are also relevant in the medical and veterinary fields, as they are pathogenic to both humans and most livestock species. Mycoplasma cells have minute sizes, often in the 300- to 800-nm range. As these dimensions are close to the diffraction limit of visible light, fluorescence imaging in mycoplasmas is often poorly informative. Recently developed superresolution imaging techniques can break this diffraction limit, improving the imaging resolution by an order of magnitude and offering a new nanoscale vision of the organization of these bacteria. These techniques have, however, not been applied to mycoplasmas before. Here, we describe an efficient and reliable protocol to perform single-molecule localization microscopy (SMLM) imaging in mycoplasmas. We provide a polyvalent transposon-based system to express the photoconvertible fluorescent protein mEos3.2, enabling photo-activated localization microscopy (PALM) in most Mycoplasma species. We also describe the application of direct stochastic optical reconstruction microscopy (dSTORM). We showcase the potential of these techniques by studying the subcellular localization of two proteins of interest. Our work highlights the benefits of state-of-the-art microscopy techniques for mycoplasmology and provides an incentive to further the development of SMLM strategies to study these organisms in the future..Dissection moléculaire du système de clivage d'anticorps des mycoplasmesExploration de Nouvelles Activités de Manipulation des Anticorp

Imaging minimal bacteria at the nanoscale: a reliable and versatile process to perform Single Molecule Localization Microscopy in mycoplasmas

Mycoplasmas are the smallest free-living organisms. These bacteria are important models for both fundamental and Synthetic Biology, owing to their highly reduced genomes. They are also relevant in the medical and veterinary fields, as they are pathogenic of both humans and most livestock species. Mycoplasma cells have minute sizes, often in the 300-800 nanometers range. As these dimensions are close to the diffraction limit of visible light, fluorescence imaging in mycoplasmas is often poorly informative. Recently developed Super-Resolution Imaging techniques can break this diffraction limit, improving the imaging resolution by an order of magnitude and offering a new nanoscale vision of the organization of these bacteria. These techniques have however not been applied to mycoplasmas before. Here, we describe an efficient and reliable protocol to perform Single-Molecule Localization Microscopy (SMLM) imaging in mycoplasmas. We provide a polyvalent transposon-based system to express the photo-convertible fluorescent protein mEos3.2, enabling Photo-Activated Localization Microscopy (PALM) in most Mycoplasma species. We also describe the application of direct STochastic Optical Reconstruction Microscopy (dSTORM). We showcase the potential of these techniques by studying the subcellular localization of two proteins of interest. Our work highlights the benefits of state-of-the-art microscopy techniques for mycoplasmology and provides an incentive to further the development SMLM strategies to study these organisms in the future

L’eau en partage

Les petits barrages constituent une composante désormais incontournable des paysages d'Afrique de l'Ouest. Petits et nombreux, mais dispersés, ils structurent les espaces, en générant un ensemble de ressources diverses. Ils induisent de fait la juxtaposition d'acteurs et d'usages souvent complémentaires, parfois concurrents, éventuellement antagonistes. Ils sont ainsi à l'origine de recompositions sociales marquées par des réarrangements institutionnels et de nouvelles règles d'accès aux écosystèmes et aux ressources. Objets de développement reconnus et appréciés des populations, leur nombre ne cesse d'augmenter. Pour évaluer les bénéfices et les risques associés à ces aménagements, cet ouvrage, centré sur les petits barrages du nord de la Côte d'Ivoire, dresse un état des lieux pluridisciplinaire. Destiné à tous les chercheurs et aménageurs préoccupés par le développement durable des savanes soudano-sahéliennes, il invite également à une réflexion plus générale sur les changements écologiques et sociaux liés à l'aménagement des vallées.Small reservoirs are now an itnportantjeature of West African landscapes. Small and numerous - but scattered - these water bodies give structure to rural areas by generating a set of varied resources. They lead to a juxtaposition of players and uses that are often complementary, sometimes in competition with each other and possibly a source of antagonism. They also induce social reconstruction marked by institutional rearrangements and new rulesfor access to ecosystems and resources. A recognised development feature appreciated by local populations, their numbers are increasing continuously. To evaluate the benefits and risks that may be associated with the presence and use of thesefacilities, this book centred on the small reservoirs in Northern Cote d'lvoireforms a multidisciplinary report on the situation. It is intended for all researchers and development bodies concerned by the sustainable development of the Sudan-Sahel savannah regions and invites broader refection on the ecological and social changes involved in the development of inland valleys