CRISPR–Cas9 genome engineering of primary CD4+ T cells for the interrogation of HIV–host factor interactions

CRISPR–Cas9 gene-editing strategies have revolutionized our ability to engineer the human genome for robust functional interrogation of complex biological processes. We have recently adapted this technology for use in primary human CD4 + T cells to create a high-throughput platform for analyzing the...

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Veröffentlicht in:	Nature protocols 2019-01, Vol.14 (1), p.1-27
Hauptverfasser:	Hultquist, Judd F., Hiatt, Joseph, Schumann, Kathrin, McGregor, Michael J., Roth, Theodore L., Haas, Paige, Doudna, Jennifer A., Marson, Alexander, Krogan, Nevan J.
Format:	Artikel
Sprache:	eng
Schlagworte:	631/208/4041 631/326/596/2557 Analytical Chemistry Antibodies - pharmacology Antigens, CD - genetics Antigens, CD - immunology Biological activity Biological Techniques Biomedical and Life Sciences Biotechnology CD4 antigen CD4-Positive T-Lymphocytes - drug effects CD4-Positive T-Lymphocytes - immunology CD4-Positive T-Lymphocytes - metabolism CD4-Positive T-Lymphocytes - virology Cell culture Cell Nucleus - drug effects Cell Nucleus - immunology Cell Nucleus - metabolism Cell Nucleus - virology Chemical synthesis Clustered Regularly Interspaced Short Palindromic Repeats Computational Biology/Bioinformatics CRISPR CRISPR-Associated Protein 9 - genetics CRISPR-Associated Protein 9 - metabolism CRISPR-Cas Systems Electroporation - methods Gene Editing - methods Genetic modification Genome editing Genome, Human Genomes High-Throughput Screening Assays HIV HIV-1 - genetics HIV-1 - immunology Host-Pathogen Interactions - genetics Host-Pathogen Interactions - immunology Human immunodeficiency virus Humans Infections Interrogation Life Sciences Lymphocyte Activation Lymphocytes Lymphocytes T Microarrays Multiplexing Organic Chemistry Pathogenesis Primary Cell Culture Proteins Protocol Questioning Ribonucleoproteins Ribonucleoproteins - genetics Ribonucleoproteins - immunology RNA, Guide, CRISPR-Cas Systems - genetics RNA, Guide, CRISPR-Cas Systems - metabolism Transfection
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