Ebola Virus Optical Pooled Screen | Secondary Screen Late Timepoint (24 hours) Images

Overview This archive contains a ~10 % curated subset of images captured 24 hours post‑infection (“late” time‑point) in HeLa and Huh7 cells with targeted CRISPR knockouts (113 genes, 6 sgRNAs per gene), read out by optical pooled screening. Full Phenotyping Image Location gs://opspublic-east1/EBOVOpticalPooledScreen/Images/SEC_Late Metadata (Cell coordinates, in‑situ sequencing reads, guide assignments) gs://opspublic-east1/EBOVOpticalPooledScreen/Metadata/Secondary Imaging Details Channel 1: DAPI – Nuclei Channel 2: VP35 protein – Ebola virus replication factories Channel 3: c‑Jun – Stress‑response transcription factor Channel 4: VP35 RNA FISH (positive‑sense) – Viral genomic/antigenomic RNA Plate Layout Row A: HeLa + EBOV | HeLa + MARV | HeLa + SUDV Row B: Huh7 + EBOV | Huh7 + MARV | Huh7 + SUDV Manuscript Context (brief) Filoviruses such as Ebola virus (EBOV) cause recurrent outbreaks with high mortality, yet therapeutic options remain scarce. Using an image‑based, genome‑wide CRISPR knockout screen spanning 39 million cells, we identified 998 host genes that modulate distinct stages of the EBOV life‑cycle. Key validated hits UQCRB – Post‑entry regulator of viral RNA synthesis; small‑molecule inhibition lowers EBOV titres in vitro. STRAP – Spliceosome‑associated factor whose disruption skews the viral RNA‑to‑protein ratio; interacts with viral co‑factor VP35. The primary screen plus 12 targeted secondary screens (including Sudan and Marburg virus assays) provide a rich resource for host dependencies and potential antiviral targets.