biostudies-arrayexpressMetabolomicsUnknownTranscriptomicsGenomicsProteomicsGeorge Kassiotisn/aIllumina NovaSeq 6000RNA-seq of coding RNAHomo sapiensHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16688H9 human embryonic stem cells (ESCs cells) were modified by CRISPR/Cas9 in order to inactivate the CALB1 gene. Wild-type (wt) and targeted (em) clones, were resset into a naive state and sequenced in triplicate.biostudies-arrayexpressSample Collection - H9 cells were grown in, stimulated and collected from culture dishesSequencing - RNA-seq libraries were sequenced on a NovaSeq 6000 (Illumina)Library Construction - Library prep was performed with NEBNext Ultra II Directional PolyA mRNA kit (NEB, Cat #E7760)Growth Protocol - H9 cells were chemically reset to naïve cells as previously described (29). DR4 MEFs, irradiated, were seeded onto 6-well plates coated with 0.1% gelatine 1-2 days before the conversion at a density of approximately 2×105 cells/well. The H9 cells were then passaged using TyplETM Select Enzyme and seeded at 5×105 cells/well in mTESRTM1 with 10 µM ROCKi for 24 h and a further 24 h in mTESTTM1 without ROCKi until medium sized colonies had formed. Chemical resetting medium was then applied at hypoxia 5% O2 for 3 days with the chemical resetting medium being replenished every 24 h. The cells were then maintained in PXGL at hypoxia, fed every day and passaged twice a week using StemPro™ Accutase™ Cell Dissociation Reagent and 10 µM ROCKi and Matrigel® hESC-Qualified Matrix for the first 24 h.Nucleic Acid Extraction - RNA from cultured H9 cells was extracted using RNeasy kit (Qiagen, Cat #74104)OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesGeorge KassiotisfalseRNA-Seq from naive CALB1wt and CALB1em H9 ESC clonesH9 human embryonic stem cells (ESCs cells) were modified by CRISPR/Cas9 in order to inactivate the CALB1 gene. Wild-type (wt) and targeted (em) clones, were resset into a naive state and sequenced in triplicate.2026-09-04T00:00:00Z2026-09-04T08:34:28.323Z2026-02-26T14:04:16.43ZE-MTAB-16688ERP189675E-MTAB-15900EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003738EFO_0004184