Keynote Presentation: Timing Is Everything: Deconstructing the Ebola Virus Infection Program by CRISPR w/ Live Q&A

Ebola virus executes its replication cycle through a tightly orchestrated sequence of events, but the host factors that govern the timing of these steps have remained poorly defined. In this talk, I will present our development and application of high-biocontainment optical pooled screening, the first large-scale, image-based CRISPR knockout screen capable of resolving the temporal program of Ebola virus infection at single-cell resolution. By integrating high-content imaging, deep learning–driven phenotypic analysis, and time-resolved infection markers, we mapped how specific host genes control discrete transitions in the viral life cycle, from entry and transcription to genome replication and egress. This approach revealed both stage-specific regulators and multifunctional factors that influence multiple phases of infection, offering new mechanistic insights and identifying potential antiviral targets. Our findings demonstrate the power of combining CRISPR screening with advanced imaging to deconvolve complex viral programs and highlight a generalizable strategy for studying other high-consequence pathogens in real time.

Learning Objectives:

1. Describe the sequence of key stages in the Ebola virus replication cycle and the importance of host factors in regulating these steps.

2. Explain how high-biocontainment optical pooled CRISPR screening combined with high-content imaging can resolve the temporal dynamics of viral infection at single-cell resolution.

3. Analyze how stage-specific and multifunctional host factors influence Ebola virus replication and identify potential antiviral targets.