PLX105188

GSE103662: iMEF SDHC-loss RNA-seq time course

• Organsim mouse
• Type RNASEQ
• Target gene
• Project ARCHS4

We have developed a tet-inducible immortalized mouse embryonic fibroblast (iMEF) cell culture model of SDH-loss paraganglioma/pheochromocytoma in which silencing gene rearrangement of the Sdhc floxed allele is driven by doxycycline-dependent expression of cre-recombinase from a tet-inducible promoter (R26M2rtTA/+;TetOcre;Sdhcfl/fl). Using this model and an isogenic Sdhc wt control line (R26M2rtTA/+;TetOcre;Sdhcfl/wt), we have characterized the time-course of Sdhc gene rearrangement, protein loss, and succinate accumulation following induction with doxycycline. Using RNA-seq, we have quantified changes in gene expression due to succinate accumulation using our Sdhc -/- cell line. Through a time-course experimental design, we have grown R26M2rtTA/+;TetOcre;Sdhcfl/fl (experimental) and R26M2rtTA/+;TetOcre;Sdhcfl/wt (control) iMEFs in standard DMEM media containing 10% FBS and quantified transcriptional changes in these cells as a function of time after triggering SDHC gene rearrangement via doxycycline exposure. SOURCE: John,Andrew,SmestadLouis (Jim) Maher Mayo Clinic