Genetic screens are powerful tools to interrogate cell response to drug treatment as well as microenvironment perturbations. With the increasing interest in manipulating the immune system to tackle disease and the use of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)–Cas9 genome editing, screening primary cells is both appealing and feasible. At Horizon we have developed a pipeline to perform pooled CRISPR screening in primary T cells, optimising a combined lentiviral and electroporation approach. We have chosen to validate our protocol using culture settings previously published in Jurkat T cells (Birsoy et al., 2015). We treated primary T cells with an electron transport chain inhibitor to identify new metabolic targets involved in cell proliferation control and survival. Our Next generation sequencing data not only confirmed some Jurkat hits, but they also reveal new specific T cell targets.
The webinar covers:
- Human T cell gene editing
- Donor variability in T cells CRISPR screening
- NGS data analysis for kinetic screenings
Dr. Cristina Ghirelli
Manager of Immuno-Oncology at Horizon Discovery