CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites

Julia K. Bialek; Thomas Walther; Joachim Hauber; Ulrike C. Lange

doi:10.3791/58572

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CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites

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免疫学と感染

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JoVE Journal 免疫学と感染

CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites

9,605 Views

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14:27 min

•

November 14, 2018

DOI:

10.3791/58572-v

DOI:

10.3791/58572-v

•

14:27 min

November 14, 2018

•

3 Views

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)

概要

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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CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites

概要

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