characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-07-07T00:23:19Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-07-07T00:23:19Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110644472
characteristics.characteristic: External Id
characteristics.text: EMBL
characteristics.characteristic: INSDC center name
characteristics.text: 2023-07-07T00:23:19Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-07-07T00:23:19Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12742238
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12101:ring1b-1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: mESC
characteristics.characteristic: cell_line
characteristics.text: embryonic stem cell
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Naïve murine embryonic stem cells (mESC) were derived freshly (FVB or mixed 129/B6)  and were routinely cultured on gelatin-coated plates in t2i/L media: NDiff (N2B27) (Takara #y40002) supplemented with titrated 2i (0.2 μM PD0325901 and 3 μM CHIR 99021), 1,000 U/ml leukaemia inhibitory factor (LIF), 1% FBS, 1% penicillin streptomycin and maintained in humidified atmosphere at 37°C and 5% CO2. Cells were passaged every 2 days by dissociation with TrypLE and medium was changed daily. Total RNA was isolated from cell pellets using NEB Monarch® Total RNA Miniprep Kit and libraries prepared using NEBNext RNA Ultra II kit (NEB E7770S, input: 350 ng of total RNA). RNA-sequencing of polyA RNA from mouse Embryonic Stem Cells. Total RNA was isolated using the Monarch® Total RNA Miniprep Kit (NEB #T2010S), including gDNA removal step and on-column DNase I digestion. Total RNA was quantified using Qubit III and quality assessed with high-sensitivity RNA Screen Tape (Agilent 5067-5579). Stranded mRNA-seq libraries were prepared with the NEBNext RNA Ultra II kit (input: 350 ng of total RNA) following the manufacturer’s instructions.
characteristics.characteristic: description
characteristics.text: stable integration of dCas9-GCN4, Ring1b-CDscFV and gRNA against Hbby locus
characteristics.characteristic: genetic modification
characteristics.text: dCas9-GCN4, Ring1b-CDscFV
characteristics.characteristic: genotype
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: FVB/NJ or mixed C57BL/6 and 129 mouse strain
characteristics.characteristic: strain
characteristics.text: ring1b-1
characteristics.characteristic: title