2024-09-13
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA115158760
External Id
Duke-NUS Medical School
INSDC center name
2024-01-26T19:40:28Z
INSDC last update
public
INSDC status
ERS17891672
SRA accession
E-MTAB-13751:Huh7.5.1 Brunello library infected with HCoV-229E
Submitter Id
ArrayExpress
broker name
Huh7.5.1
cell_line
not collected
collection date
human
common name
Protocols: For DENV and JEV, a pool of ~40 million mutagenized cells – about 500-fold coverage for each unique sgRNA from the library – were used for each of the screen. The seeded mutagenized cells were subsequently infected with DENV or JEV at a MOI of 0.2 and 0.05 respectively the next day. An additional round of infection was carried out at 18 days post infection. Resistant cells were harvested in two batches for each screen, one at 21 days post infection and the other at 27 days post infection. Cells harvested in the second batch were also reinfected at 21 days post initial infection. A total of 44 million non-infected mutagenized cells were simultaneously harvested to serve as the unselected control. For HCoV-229E screen, a pool of ~60 million mutagenized cells were subsequently seeded and infected with HCoV-229E at a MOI of 0.003. Resistant cells were harvested at 15 days post infection. A total number of 40 million non-infected populations of mutagenized cells were harvested in parallel as the unselected control. Genomic DNA were extracted from each harvested cell pellet using QIAamp DNA Mini Kit (Qiagen). The sgRNA containing sequence were PCR amplified and barcoded with Illumina next generation sequencing indexes.
description
CRISPR perturbed mutant library
genotype
not collected
geographic location (country and/or sea)
HCoV-229E
infect
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
#1
replicate
Homo sapiens
scientific_name
Huh7.5.1 Brunello library infected with HCoV-229E
title