title: RO7
organism: Homo sapiens
description: 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.).
attributes.tag: ENA first public
attributes.value: 2025-04-30
attributes.tag: INSDC center name
attributes.value: Division of Cancer Sciences, Faculty of Biology, Medicine and Health, University of Manchester
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-14654:RO7
attributes.tag: batch
attributes.value: 1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_line
attributes.value: RKO
attributes.tag: cell_type
attributes.value: colorectal
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: colon carcinoma
attributes.tag: genotype
attributes.value: Tet_Inducible
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: myc
attributes.value: High
attributes.tag: repeat
attributes.value: B
attributes.tag: sample name
attributes.value: E-MTAB-14654:RO7
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: tet
attributes.value: Yes