characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-08-09T00:19:20Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-08-09T00:19:20Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112328711
characteristics.characteristic: External Id
characteristics.text: Hebrew University, Genetics Department
characteristics.characteristic: INSDC center name
characteristics.text: 2023-08-09T00:19:20Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-08-09T00:19:20Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14439626
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12505:SARS_Rep1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: h-pES10
characteristics.characteristic: cell_line
characteristics.text: endoderm cell
characteristics.characteristic: cell_type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: Cell culture and maintainance: For the genetic screen the haploid hESC line, h-pES10, was used to generate a genome-wide loss-of-function library. The mutant haploid hESC library was cultured at 37 °C and 5% CO2 in feeder-free conditions on matrigel-coated plates (Corning) in mTeSR1 (STEMCELL Technologies) supplemented with 10 μM ROCK inhibitor Y-27632 (Stemgent) for 1 day after splitting. Before reaching confluency, cells were passaged by a quick trypsinization using TrypLE Select (Thermo Fisher Scientific). Vero E6 cells were maintained in Eagle's Minimum Essential Medium (EMEM; Biological Industries, Beit Haemek, Israel), supplemented with 10% fetal bovine serum, 2 mM L-Glutamine, 10 IU/ml Penicillin, and 10 μg/ml streptomycin (Biological Industries, Beit Haemek, Israel). Definitive endoderm differentiation assay: Mutant h-pES10 library cells were used for differentiation into definitive endoderm (DE). Cells were plated on Matrigel-coated 6-well plates at a density of 2 million cells per well. The next day, STEMdiff™ Definitive Endoderm Kit (STEMCELL Technologies, cat. no. 05110) has been used according to manufacturer’s instructions. Briefly, the differentiation medium (2 ml/well) with its appropriate supplements was replaced every 24 hours for 4 days. SARS-CoV-2 Screen: On the 5th day of differentiation, differentiated DE cells were infected with SARS-CoV-2 at various MOIs. Following infection, cells were continued to be cultured in the DE medium for additional 5 days. Cells were allowed to undergo viral cytotoxicity and harvested 5 days post-infection for further analysis. 2 screen samples were infected with SARS-COV-2, and one control screen sample was not infected. Samples were harvested using trypsin gSYNC DNA extraction kit (Geneaid) PCR with Nextera overhangs
characteristics.characteristic: description
characteristics.text: embryo
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: CRISPR/Cas9 loss of function library
characteristics.characteristic: genotype
characteristics.text: Severe acute respiratory syndrome coronavirus 2
characteristics.characteristic: infect
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: inner cell mass
characteristics.characteristic: organism part
characteristics.text: embryonic stem cell
characteristics.characteristic: progenitor cell type
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: SARS_Rep1
characteristics.characteristic: title