2025-12-31
ENA first public
ERC000011
ENA-CHECKLIST
Duke-NUS Medical School
INSDC center name
public
INSDC status
ERS21101800
SRA accession
E-MTAB-14485:Pteropine Orthoreovirus PRV3M
Submitter Id
ArrayExpress
broker name
IGROV-1
cell_line
Ovarian endometrioid adenocarcinoma
cell_type
not collected
collection date
human
common name
Protocols: 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. 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 instructions 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.
description
adult
developmental stage
Endometrioid carcinoma of ovary
disease
genome-wide CRISPR KO library under phenotypic selection
genotype
not collected
geographic location (country and/or sea)
Orthoreovirus PRV3M
infect
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
ovary
organism part
Homo sapiens
scientific_name
Pteropine Orthoreovirus PRV3M
title