Poster: A Genome-Wide CROP-Seq Screen Reveals Mediators of T Cell Signaling

The first-of-its-kind genome-scale CRISPRi screen was conducted to verify factors involved in TCR signaling pathways. Unravelling these T cell-intrinsic molecular factors could potentially unlock new therapeutic targets and strategies for modulating T cell responses in various immune-related disorders.

In more detail, a guide RNA library targeting 18,595 human genes was utilized for CRISPR-based gene knockdowns in Jurkat cells expressing the dCas9-KRAB fusion endonuclease. In total, one million Jurkat cells were processed for single-cell RNA sequencing allowing transcriptomic readouts of a final list of 374 marker genes involved in TCR signaling.

The bioinformatic analysis confirmed more than 70 known activators and repressors of TCR signaling cascades, hence showcasing the potential of Perturb-seq (CROP-seq) screens to support translational research.

Recently, the first-of-its-kind genome-scale CRISPRi screen was featured in a 10x Genomics webinar and you can find the video in the “How it works” section:

Submit your credentials and download the corresponding poster!