biostudies-arrayexpressYaw Shin OoiHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-14485Unbiased forward genetic screen to identify host factors for Pteropine Orthoreovirus PRV3Mbiostudies-arrayexpressSample Collection - A pool of mutagenized IGROV-1 cells were generated using the Brunello CRISPR knockout library, which was designed to target human genome with four unique sgRNA per gene. The mutagenized IGROV-1 cells were subjected to pteropine orthoreovirus PRV3M infection. After a total of two weeks of live/dead selection, mutagenized cells resisting cytolytic infection of pteropine orthoreovirus PRV3M was harvested fand subjected to NGS analysis.Nucleic Acid Extraction - Resistant cells were harvested from each sub-library, and subjected to total genomic DNA extraction using DNA mini kit (Qiagen) according to the manufacturer’s instructionsSequencing - Illumina MiSeq (50bp run)Library Construction - The sgRNA were amplified from gDNA in two-step PCR using Herculase II Fusion DNA Polymerase (Agilent). For first PCR, 20 ug of gDNA for each library was amplified for 16 cycles. For 2nd PCR, 1 reaction containing 5ul PCR product for each sub-library was amplified for 27 cycles, using Illumina indexed primers. PCR products were gel purified (Qiagen gel purification kit) and sequenced on Illumina MiSeq platform.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - NGS data was analyzed using MAGeCK.UnknownTranscriptomicsGenomicsProteomicsIllumina MiSeqBSL2 LaboratoryDNA-seqHomo sapiensYaw Shin OoifalseGenome-scale CRISPR knockout screen for Pteropine Orthoreovirus PRV3MUnbiased forward genetic screen to identify host factors for Pteropine Orthoreovirus PRV3M2025-12-31T00:00:00Z2025-12-31T02:02:12.854Z2024-10-01T10:13:58.515ZE-MTAB-14485ERP164647EFO_0002944EFO_0004170EFO_0002693EFO_0005518EFO_0003816EFO_0004184