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Heinrich-Pette-Institute for Experimental Virology and Immunology, University of Hamburg

2 ARTICLES PUBLISHED IN JoVE

Biology

Interview: HIV-1 Proviral DNA Excision Using an Evolved Recombinase

¹Department of Cell Biology and Virology, Heinrich-Pette-Institute for Experimental Virology and Immunology, University of Hamburg

Current HIV-1 strategies act to suppress the viral life cycle but do not effectively eradicate infection. Here, we demonstrate that an engineered recombinase can efficiently excise integrated HIV-1 proviral DNA from the genome of infected cells.

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.