SEP 18, 2018 8:00 AM PDT

High-throughput CRISPR-Cas9 Genome Engineering in Primary T Cells

Sponsored by: Horizon Discovery
C.E. Credits: P.A.C.E. CE Florida CE
  • Assistant Professor of Medicine, Northwestern University
      Judd F. Hultquist, Ph.D., is an Assistant Professor of Medicine at Northwestern University in Chicago, Illinois. His lab specializes in the adaptation of proteomic and functional genomic technologies to primary models of disease to better understand the molecular dynamics of the host-pathogen relationship. His work on CRISPR technologies with Dr. Nevan Krogan at the University of California, San Francisco led to the development of the first gene editing platform for primary CD4+ T cells, the identification of several novel pathways that contribute to HIV replication, and the discovery of a new class of CRISPR inhibitors encoded by bacteriophages that maintain their function in mammalian cells. He is currently funded by the Third Coast Center for AIDS Research (CFAR) and the American Foundation for AIDS Research (amFAR) to apply CRISPR-Cas9 RNP technology to uncover the molecular mechanisms that underlie HIV latency and reactivation in patient reservoirs.

    DATE: September 18, 2018
    TIME: 08:00am PDT, 11:00am EDT
    CRISPR-Cas9 gene editing strategies have revolutionized our ability to engineer the genomes of diverse cell types and species for the robust functional interrogation of complex biological processes. Gene editing in primary T-cells represents an important research tool for the development of T-cell based immunotherapies, the study of the autoimmunity, the understanding of host-pathogen dynamics that drive HIV replication and latency, and creation of chimeric antigen receptor (CAR) T cells for the treatment of various cancers. While these cells have proven resistant to traditional transfection and transduction approaches for the CRISPR-Cas9 gene editing, recent advancements in ribonucleoprotein (RNP) delivery have allowed for efficient primary T-cell editing for the first time. In this webinar, we will present a workflow for the design and delivery of CRISPR-Cas9 RNPs to primary, human T-cells by electroporation. This platform supports the high-throughput, arrayed generation of hundreds of specific gene manipulations in only a few hours time without the use of recombinant DNA or lentiviruses and without the need for selection markers. This approach is widely adaptable to an array of downstream applications and protocols for the study of diverse T-cell processes.
    In this webinar, you will learn:
    • Optimal conditions for CRISPR-Cas9 gene editing in primary T cells using RNPs
    • Rules for designing, synthesizing, and validating efficient guide RNAs
    • Considerations for high-throughput functional screening
    • How this approach is being employed to better understand HIV replication and pathogenesis


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