IMEF SDHC-loss stable line RNA-seq

ABSTRACT: We have developed a tet-inducible SV-40 lentivirus-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). For comparison, we include an isogenic Sdhc wt control line (R26M2rtTA/+;TetOcre;Sdhcfl/wt) that retains one intact copy of Sdhc upon doxycycline exposure. Following doxycycline induction of both cell lines, dilutional subcloning was employed to obtain stable Sdhc -/- (experimental) and Sdhc +/- (control) cell lines. Sdhc gene rearrangement status for each clonally-derived cell line was confirmed by PCR. Cell lines were grown in standard DMEM containing penicillin/streptomycin antibiotics (0.5 mg/mL), non-essential amino acids (100 micromolar each of glycine, alanine, asparagine, aspartic acid, glutamic acid, proline, and serine), sodium pyruvate (1 mM), and HEPES buffer (10 mM) at 21% O2 and 5% CO2. Following cell line derivation and verification of Sdhc genetic status, transcriptomic profiling for each cell line was performed by RNA-seq.

ORGANISM(S): Mus musculus

PROVIDER: GSE114244 | GEO | 2018/05/10

REPOSITORIES: GEO