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Leibniz Institute for Experimental Virology

3 ARTICLES PUBLISHED IN JoVE

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Biochemistry

Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis
Kathrin Rösch 1, Marcel Kwiatkowski 2, Hartmut Schlüter 2, Eva Herker 1
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.

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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 Hauber 1,3, Ulrike C. Lange 1,2,3
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.

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Biology

Chromatin Extraction from Frozen Chimeric Liver Tissue for Chromatin Immunoprecipitation Analysis
Andrea Pirosu 1,3, Lena Allweiss 1, Maura Dandri 1,2
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).

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