NGS-13069
Homo sapiens
Protocols: HepG2 cells were cultured in Minimum Essential Media (MEM) with 10% FBS. All media was supplemented with 2mM L-glutamine, 100U/ml penicillin, and 100μg/ml streptomycin. sgRNA pools are described in the additional files associated with the submission Table_S1_sgrna_metadata.tsv and Table_S6_pools.tsv. HepG2 cells inducible for Cas9 expression were generated as previously described (Henser-Brownhill et al., 2017; 10.1080/15592294.2017.1395121). Cas9 induction was induced with 1ug/mL doxycycline 1 day prior to infection with the sgRNAs and sustained until cells were harvested for genomic DNA extraction. Transduced cells were selected with 4ug/mL blasticidin, starting 24h after the first infection, and genomic DNA was extracted 5 days after infection (Qiagen 69506). DNA libraries enriched for the targeted sites were generated using the SureSelect Target enrichment kit (Agilent) following the manufacturer's instructions. Capture probes were designed to cover 2Kb regions centered on each target site.
ENA-FIRST-PUBLIC
2018-12-13T17:03:14Z
ENA-LAST-UPDATE
2018-07-31T14:11:02Z
External Id
SAMEA4810434
INSDC center name
The Francis Crick Institute
INSDC first public
2018-12-13T17:03:14Z
INSDC last update
2018-07-31T14:11:02Z
INSDC status
public
Submitter Id
E-MTAB-7095:NGS-13069
broker name
ArrayExpress
cell line
HepG2
cell type
epithelial cell
common name
human
disease
hepatocellular carcinoma
genotype
CRISPR/Cas9 mediated knockout with sgRNA pool 450-5
organism part
liver
sample name
E-MTAB-7095:NGS-13069
scientific_name
Homo sapiens