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Immunology and Infection

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

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

Article DOI: 10.3791/58572-v • 14:27 min • November 14th, 2018
November 14th, 2018 •
*1,2, *1, 1,3, 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)
* These authors contributed equally

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résumé
Automatic Translation

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.

Tags

CRISPR-Cas9-based Genome Engineering Jurkat Reporter Models HIV-1 Infection Proviral Integration Sites HIV Provirus Integration Androgynous Gene Expression Genomic Integration Sites In-vitro Models Pro-viral Reporters CRISPR/Cas9-based Genome Engineering UCSC Genome Browser In-silico-extract Genomic Sequence Guide RNAs E-CRISP Web Tool Homo Sapiens Genome Reference Consortium Human Genome:Build 38.Reference Genome Guide-RNA Search PAM Off-targets Tolerate Mismatches Excluded Introns CPG Islands NCBI Blast Browser
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