CRISPRa Perturb-seq decodes stimulation responses in primary human T cells

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. University of California San Francisco researchers 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.

Genome-wide CRISPRa/i screens discover tunable regulators of stimulation-responsive cytokine production in primary human T cells. Genome-wide CRISPRa/i gain-of-function and loss-of-function screens in human T cells allowed for systematic identification of regulators of cytokine production. Follow-up on key CRISPRa screen hits with secretome and scRNA-seq analysis helped to decode how these regulators tune T cell activation and program cells into different stimulation-responsive states.

Schmidt R, Steinhart Z, Layeghi M, Freimer JW, Bueno R, Nguyen VQ, Blaeschke F, Ye CJ, Marson A. (2022) CRISPR activation and interference screens decode stimulation responses in primary human T cells. Science 375(6580):eabj4008. [article]

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