MAY 21, 2019 7:00 AM PDT

Genome-wide CRISPR Screens in Primary Human T Cells Reveal Key Regulators of Immune Function

  • Postdoctoral Fellow, Department of Microbiology and Immunology, University of California, San Francisco
      Eric Shifrut is a postdoctoral fellow in Alexander Marson's group at the University of California, San Francisco. Eric completed his PhD at the Weizmann Institute of Science in Israel, under supervision of Prof. Nir Friedman. In his thesis work, Eric developed computational and experimental platforms to map the landscape of T cell receptor repertoires in health and disease. Since joining the Marson lab in 2016, Eric has been working to pioneer CRISPR applications in primary human T cells. These efforts aim to design discovery platforms to explore and exploit genetic circuits to boost T cell function in cancer immunotherapy.

    DATE:  May 21, 2019
    TIMEL  7:00am PT, 10:00am ET
    Human T cells are central effectors of immunity and cancer immunotherapy. CRISPR-based functional studies in T cells could prioritize novel targets for drug development and improve the design of genetically reprogrammed cell-based therapies. However, large-scale CRISPR screens have been challenging in primary human cells. Critical biology of human immune cells, including key signaling pathways and effector functions, may not be recapitulated in immortalized cell lines. We developed a new method, sgRNA lentiviral infection with Cas9 protein electroporation (SLICE), to identify regulators of stimulation responses in primary human T cells. Genome-wide loss-of-function screens identified essential T cell receptor signaling components and genes that negatively tune proliferation following stimulation. Targeted ablation of individual candidate genes characterized hits and identified perturbations that enhanced cancer cell killing. SLICE coupled with single-cell RNA-Seq revealed signature stimulation-response gene programs altered by key genetic perturbations. SLICE genome-wide screening was also adaptable to identify mediators of immunosuppression, revealing genes controlling responses to adenosine signaling. In summary, we have developed a novel pooled CRISPR screening technology with the potential to explore unmapped genetic circuits in primary human cells and to guide the design of engineered cell therapies.
    Learning Objectives:
    • Identify the components of a new pooled CRISPR screening technique (SLICE) using molecular biology
    • Understand how to explore unmapped genetic circuits in primary human cells using SLICE
    • Learn how to design engineered cell therapies using SLICE
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    MAY 21, 2019 7:00 AM PDT

    Genome-wide CRISPR Screens in Primary Human T Cells Reveal Key Regulators of Immune Function


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