Leibniz Institute for Experimental Virology 4 articles published in JoVE Biology Chromatin Extraction from Frozen Chimeric Liver Tissue for Chromatin Immunoprecipitation Analysis Andrea Pirosu1,3, Lena Allweiss1, Maura Dandri1,2 1Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, 2German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel-Riems site, 3Research Department Virus Immunology, Leibniz Institute for Experimental Virology This protocol focuses on chromatin preparation from snap frozen tissues and it is suitable for Crosslinking Chromatin Immunoprecipitation (X-ChIP) followed by either quantitative PCR analysis (X-ChIP-qPCR) or next generation sequencing approaches (X-ChIP-seq). Immunology and Infection CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites Julia K. Bialek*1,2, Thomas Walther*1, Joachim Hauber1,3, Ulrike C. Lange1,2,3 1Heinrich Pette Institute, Leibniz Institute for Experimental Virology, 2Department of Anesthesiology, University Medical Center Hamburg-Eppendorf, 3German Center for Infection Research (DZIF) We present a genome engineering workflow for the generation of new in vitro models for HIV-1 infection that recapitulate proviral integration at selected genomic sites. Targeting of HIV-derived reporters is facilitated by CRISPR-Cas9-mediated, site-specific genome manipulation. Detailed protocols for single-cell clone generation, screening, and correct targeting verification are provided. Biochemistry Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis Kathrin Rösch1, Marcel Kwiatkowski2, Hartmut Schlüter2, Eva Herker1 1Heinrich Pette Institute, Leibniz Institute for Experimental Virology, 2Core Facility Mass Spectrometric Proteomics, University Medical Center Hamburg-Eppendorf Lipid droplets are important organelles for the replication of several pathogens, including the Hepatitis C Virus (HCV). We describe a method to isolate lipid droplets for quantitative mass spectrometry of associated proteins; it can be used under a variety of conditions, such as virus infection, environmental stress, or drug treatment. Immunology and Infection Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity José Vicente Pérez-Girón1, Sergio Gómez-Medina1, Anja Lüdtke1, Cesar Munoz-Fontela1 1Laboratory of Emerging Viruses, Heinrich Pette Institute, Leibniz Institute for Experimental Virology There is an overall lack of knowledge about how vaccines work. Here we propose the combined use of reverse genetics and bone marrow chimeric mice to gain insight into the early host immune responses to vaccines with a special focus on dendritic cells and T cell immunity.
