characteristics.text: 2025-04-30
characteristics.characteristic: ENA first public
characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: Division of Cancer Sciences, Faculty of Biology, Medicine and Health, University of Manchester
characteristics.characteristic: INSDC center name
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS22467424
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-14654:RO9
characteristics.characteristic: Submitter Id
characteristics.text: 1
characteristics.characteristic: batch
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: RKO
characteristics.characteristic: cell_line
characteristics.text: colorectal
characteristics.characteristic: cell_type
characteristics.text: not collected
characteristics.characteristic: collection date
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: Human colon carcinoma cell line RKO (ATCC; Cat#CRL-2577) was used. Both endogenous MYC alleles were mutated using CRISPR/Cas9-mediated gene editing. Then, Flp-mediated recombination was used to insert a tetracycline-responsive MYC transgene into a pre-existing FRT site, thus generating CRISPR-Flp-MYC cells RKO cells were cultured in Dulbecco's Modified Eagle Medium (DMEM). RKO Flp-InTM T-RexTM FC7-MYC cells were cultured in DMEM with 100 ng/ml tetracycline, 8 μg/ml blasticidin and 400 μg/ml hygromycin B (Sigma Aldrich), as described previously (Littler et al., 2019).  RKO Flp-InTM T-RexTM FC-MYC cells were seeded into T75 flasks at 300,000 (MYC-high) or 400,000 (MYC-low) cells/flask with either DMSO or 25 nM SAEi ± 500 ng/mL tetracycline. After 24 hours, cells were scraped into the media, centrifuged and resuspended in PBS, pelleted and stored at -80°C before RNA extraction RNA extraction was performed using RNeasy Micro Kit (Qiagen) following manufacturer’s instructions. RNA was quantified using a NanoDrop One (Thermo Scientific) and 1 μg stored at -80°C Libraries were generated using the TruSeq Stranded mRNA assay (Illumina, Inc.) following manufacturer’s instructions. Adaptor indices were used to allow multiplexing of pooled libraries before cluster generation using a cBot instrument (Illumina, Inc.).
characteristics.characteristic: description
characteristics.text: colon carcinoma
characteristics.characteristic: disease
characteristics.text: Tet_Inducible
characteristics.characteristic: genotype
characteristics.text: not collected
characteristics.characteristic: geographic location (country and/or sea)
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Low
characteristics.characteristic: myc
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: C
characteristics.characteristic: repeat
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: No
characteristics.characteristic: tet
characteristics.text: RO9
characteristics.characteristic: title