Optical Pooled CRISPR Screen Reveals Regulators of NF-κB Dynamics in Human Cells

Integrated pooled CRISPR screening linked to imaging readouts accelerate target identification and functional characterization of signaling pathways. A good example of this can be found in studies of NF-κB signaling, which is central to inflammatory responses and driven by rapid nuclear translocation of the p50/p65 complex to activate transcriptional programs following cytokine stimulation.

In this GEN webinar, Tilmann Buerckstuemmer, PhD, CSO at Myllia Biotechnology will show how high-throughput pooled CRISPR screening combined with cell painting readouts characterized important signaling pathways using NF-κB nuclear translocation as a case study. During the webinar, you will learn how the AVITI24 platform from Element Biosciences profiled ~440,000 cells in a pooled CRISPR screen targeting 195 genes. Linking genetic perturbations to p65 subcellular localization and cell painting features in a single workflow enabled identification of known pathway components, uncovered regulatory roles for chromatin-modifying complexes, and improved interpretation of phenotypic outcomes using morphological features.

Key takeaways include:

• Strategies for linking CRISPR perturbations to protein localization and morphological features at single-cell resolution
• Identification of hitherto poorly characterized chromatin modifying complexes in regulating NF-κB signaling
• The value of multimodal readouts, including morphology, in adding depth and confidence to recovered biology
• How this approach supports mechanism-of-action studies and enables identification of both positive and negative regulators of signaling pathways

A live Q&A session will follow the presentation offering you a chance to pose questions to our expert panelists.

Produced with support from: