Assistant Professor of Medicine, Northwestern University
BIOGRAPHY
SEP 18, 2018 8:00 AM PDT
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High-throughput CRISPR-Cas9 Genome Engineering in Primary T Cells
Speaker
Abstract
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:
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Optimal conditions for CRISPR-Cas9 gene editing in primary T cells using RNPs
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Rules for designing, synthesizing, and validating efficient guide RNAs
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Considerations for high-throughput functional screening
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How this approach is being employed to better understand HIV replication and pathogenesis
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