Leibniz Institute for Experimental Virology

3 ARTICLES PUBLISHED IN JoVE

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

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.

Biology

Chromatin Extraction from Frozen Chimeric Liver Tissue for Chromatin Immunoprecipitation Analysis

Andrea Pirosu^1,3, Lena Allweiss¹, 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).