Статья

CRISPR activation and interference screens decode stimulation responses in primary human T cells

Ralf SchmidtDepartment of Medicine, University of California San Francisco, San Francisco, CA 94143, USAZachary SteinhartDepartment of Medicine, University of California San Francisco, San Francisco, CA 94143, USAMadeline LayeghiGladstone-UCSF Institute of Genomic Immunology, San Francisco, CA 94158, USAJacob W. FreimerDepartment of Genetics, Stanford University, Stanford, CA 94305, USARaymund BuenoDepartment of Medicine, University of California San Francisco, San Francisco, CA 94143, USAVinh NguyenDepartment of Surgery, University of California San Francisco, San Francisco, CA 94143, USAFranziska BlaeschkeDepartment of Medicine, University of California San Francisco, San Francisco, CA 94143, USAChun YeChan Zuckerberg Biohub, San Francisco, CA 94158, USAAlexander MarsonChan Zuckerberg Biohub, San Francisco, CA 94158, USA

2022en

ABI

Аннотация

Regulation of cytokine production in stimulated T cells can be disrupted in autoimmunity, immunodeficiencies, and cancer. Systematic discovery of stimulation-dependent cytokine regulators requires both loss-of-function and gain-of-function studies, which have been challenging in primary human cells. We now report genome-wide CRISPR activation (CRISPRa) and interference (CRISPRi) screens in primary human T cells to identify gene networks controlling interleukin-2 (IL-2) and interferon-γ (IFN-γ) production. Arrayed CRISPRa confirmed key hits and enabled multiplexed secretome characterization, revealing reshaped cytokine responses. Coupling CRISPRa screening with single-cell RNA sequencing enabled deep molecular characterization of screen hits, revealing how perturbations tuned T cell activation and promoted cell states characterized by distinct cytokine expression profiles. These screens reveal genes that reprogram critical immune cell functions, which could inform the design of immunotherapies.

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Идентификаторы

DOI: 10.1126/science.abj4008

Цитирования и источники

Цитирований: 2Использованных источников: 0

Показатели — AkademScholar