characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2020-05-12T04:07:20Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2019-10-23T14:38:21Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA6109400
characteristics.characteristic: External Id
characteristics.text: Max Planck Institute for Infection Biology, Department of Cellular Microbiology
characteristics.characteristic: INSDC center name
characteristics.text: 2020-05-12T04:07:20Z
characteristics.characteristic: INSDC first public
characteristics.text: 2019-10-23T14:38:21Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS3910074
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-8459:Sample 131
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: PLB-985
characteristics.characteristic: cell line
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: PLB-985 at day 0, day 3 and day 7 of differentiation (see growth protocol) were collected, total RNA was extracted by lysis in well. PLB-985 were cultured in RPMI (RPMI 1640 medium, gibco) supplemented with 10 % FCS, L-glutamine and antibiotics (25030-024 and 15140-122, gibco), cells were passaged every 3-4 days. For differentiation, cells were counted at d0 and 0.4x106 cells/ml were seeded in differentiation medium (RPMI supplemented with 2.5 % FCS, 0.5% dimethylformamide (DMF, D4551, Sigma), L-glutamine and antibiotics), usually in 6 well plates in a total volume of 3ml. At d4 of differentiation, 2ml of differentiation medium were added per 6 well. PLB-985 were transduced with lentiviral particles containing the sgRNA of interest cloned into lentiCRISPRv2, cells were selected with puromycin (2.5 g/ml) and, if single clones were analyzed, seeded at a density of 0.8 cells/well in 96 wells for limited dilution cloning. Clones were sequenced using the OutKnocker protocol and software24 (Supplementary Table 3). Clones with out of frame indels leading to efficient gene disruption were used for subsequent experiments. Total RNA of PLB-985 at various time points was isolated using RNeasy kit (Qiagen) according to manual. A stranded RNA seq library was constructed using NEB-Next Ultra RNA directional kit with Poly-A tail enrichment.
characteristics.characteristic: description
characteristics.text: adult
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001272
characteristics.characteristic: developmental stage
characteristics.text: acute myeloid leukemia
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0000222
characteristics.characteristic: disease
characteristics.text: H1.2 knockout clone2
characteristics.characteristic: genotype
characteristics.text: neutrophil differentiation
characteristics.characteristic: growth condition
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: blood
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000178
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: Sample 131
characteristics.characteristic: title