DOI: 10.7910/dvn/6fqnua
title: Ebola Virus Optical Pooled Screen | Secondary Screen Early Timepoint (16 hours) Images

description: &lt;!DOCTYPE html&gt;
&lt;html lang="en"&gt;
&lt;head&gt;
&lt;meta charset="utf-8"&gt;

&lt;style&gt;
body { font-family: Arial, Helvetica, sans-serif; line-height: 1.55; margin: 24px auto; max-width: 960px; }
code, pre { background: #f4f4f4; padding: 4px 6px; border-radius: 4px; display: block; overflow-x: auto; }
table { border-collapse: collapse; margin: 14px 0; width: 100%; }
th, td { border: 1px solid #ccc; padding: 8px 12px; text-align: left; vertical-align: top; }
thead th { background: #fafafa; }
h1 { font-size: 1.9em; margin-bottom: 0.4em; }
h2 { font-size: 1.45em; margin-top: 1.3em; }
&lt;/style&gt;

&lt;/head&gt;
&lt;body&gt;

&lt;h2&gt;Overview&lt;/h2&gt;
&lt;p&gt;This archive contains a &lt;strong&gt;~10 % curated subset&lt;/strong&gt; of images captured &lt;strong&gt;16 hours post‑infection (“early” time‑point)&lt;/strong&gt; in &lt;strong&gt;HeLa&lt;/strong&gt; and &lt;strong&gt;Huh7&lt;/strong&gt; cells with targeted CRISPR knockouts (113 genes, 6 sgRNAs per gene), read out by &lt;em&gt;optical pooled screening&lt;/em&gt;.&lt;/p&gt;

&lt;h2&gt;Full Phenotyping Image Location&lt;/h2&gt;
&lt;pre&gt;&lt;code&gt;gs://opspublic-east1/EBOVOpticalPooledScreen/Images/SEC_Early&lt;/code&gt;&lt;/pre&gt;

&lt;h2&gt;Metadata&lt;/h2&gt;
&lt;p&gt;(Cell coordinates, in‑situ sequencing reads, guide assignments)&lt;/p&gt;
&lt;pre&gt;&lt;code&gt;gs://opspublic-east1/EBOVOpticalPooledScreen/Metadata/Secondary&lt;/code&gt;&lt;/pre&gt;

&lt;h2&gt;Imaging Details&lt;/h2&gt;
&lt;table&gt;
&lt;thead&gt;
&lt;tr&gt;&lt;th&gt;Channel No.&lt;/th&gt;&lt;th&gt;Marker / Stain&lt;/th&gt;&lt;th&gt;Description&lt;/th&gt;&lt;/tr&gt;
&lt;/thead&gt;
&lt;tbody&gt;
&lt;tr&gt;&lt;td&gt;1&lt;/td&gt;&lt;td&gt;&lt;strong&gt;DAPI&lt;/strong&gt;&lt;/td&gt;&lt;td&gt;Nuclei&lt;/td&gt;&lt;/tr&gt;
&lt;tr&gt;&lt;td&gt;2&lt;/td&gt;&lt;td&gt;&lt;strong&gt;VP35 protein&lt;/strong&gt;&lt;/td&gt;&lt;td&gt;Ebola virus replication factories&lt;/td&gt;&lt;/tr&gt;
&lt;tr&gt;&lt;td&gt;3&lt;/td&gt;&lt;td&gt;&lt;strong&gt;c‑Jun&lt;/strong&gt;&lt;/td&gt;&lt;td&gt;Stress‑response transcription factor&lt;/td&gt;&lt;/tr&gt;
&lt;tr&gt;&lt;td&gt;4&lt;/td&gt;&lt;td&gt;&lt;strong&gt;VP35 RNA FISH&lt;/strong&gt; (positive‑sense)&lt;/td&gt;&lt;td&gt;Viral genomic / antigenomic RNA&lt;/td&gt;&lt;/tr&gt;
&lt;/tbody&gt;
&lt;/table&gt;

&lt;h2&gt;Plate Layout&lt;/h2&gt;
&lt;table&gt;
&lt;thead&gt;
&lt;tr&gt;&lt;th&gt;Row&lt;/th&gt;&lt;th&gt;Col 1 (A1/B1)&lt;/th&gt;&lt;th&gt;Col 2 (A2/B2)&lt;/th&gt;&lt;th&gt;Col 3 (A3/B3)&lt;/th&gt;&lt;/tr&gt;
&lt;/thead&gt;
&lt;tbody&gt;
&lt;tr&gt;&lt;td&gt;A&lt;/td&gt;&lt;td&gt;HeLa + EBOV&lt;/td&gt;&lt;td&gt;HeLa + MARV&lt;/td&gt;&lt;td&gt;HeLa + SUDV&lt;/td&gt;&lt;/tr&gt;
&lt;tr&gt;&lt;td&gt;B&lt;/td&gt;&lt;td&gt;Huh7 + EBOV&lt;/td&gt;&lt;td&gt;Huh7 + MARV&lt;/td&gt;&lt;td&gt;Huh7 + SUDV&lt;/td&gt;&lt;/tr&gt;
&lt;/tbody&gt;
&lt;/table&gt;
&lt;p&gt;(Three technical replicates per condition.)&lt;/p&gt;

&lt;h2&gt;Manuscript Context (brief)&lt;/h2&gt;
&lt;p&gt;Filoviruses such as &lt;strong&gt;Ebola virus (EBOV)&lt;/strong&gt; cause recurrent outbreaks with high mortality, yet therapeutic options remain scarce. Using an &lt;strong&gt;image‑based, genome‑wide CRISPR knockout screen&lt;/strong&gt; spanning &lt;strong&gt;39 million cells&lt;/strong&gt;, we identified &lt;strong&gt;998 host genes&lt;/strong&gt; that modulate distinct stages of the EBOV life‑cycle.&lt;/p&gt;

&lt;p&gt;&lt;strong&gt;Key validated hits&lt;/strong&gt;&lt;/p&gt;
&lt;ul&gt;
&lt;li&gt;&lt;strong&gt;UQCRB&lt;/strong&gt; – post‑entry regulator of viral RNA synthesis; small‑molecule inhibition lowers EBOV titres &lt;em&gt;in vitro&lt;/em&gt;.&lt;/li&gt;
&lt;li&gt;&lt;strong&gt;STRAP&lt;/strong&gt; – spliceosome‑associated factor whose disruption skews the viral RNA‑to‑protein ratio; interacts with viral co‑factor &lt;strong&gt;VP35&lt;/strong&gt;.&lt;/li&gt;
&lt;/ul&gt;

&lt;p&gt;The primary screen plus &lt;strong&gt;12 targeted secondary screens&lt;/strong&gt; (including Sudan and Marburg virus assays) provide a rich resource for host dependencies and potential antiviral targets.&lt;/p&gt;

&lt;p&gt;For full imaging parameters, staining protocols, and biosafety procedures, consult the manuscript.&lt;/p&gt;

&lt;/body&gt;
&lt;/html&gt;