Cytochrome c Stable Knockout Mouse Fibroblast Cell Line

Mice harboring a heterozygous somatic cyt c KO allele (cyt cs⁺ᐟ⁻) with mice harboring a homozygous testis cyt c KO allele (cyt ct⁻ᐟ⁻) were crossed. We used homozygous cyt ct⁻ᐟ⁻ females for the crosses. Embryonic lethal phenotype was rescued by introducing a ubiquitously expressed somatic cyt c transgene flanked by loxP sites. Crosses of one of the transgenic animals with the cyt cyt⁻ᐟ⁻cyt cs⁺ᐟ⁻ animals gave rise to F1 mice that when backcrossed produced mice with the cyt ct⁻ᐟ⁻cyt cs⁻ᐟ⁻- cyt cflox/o genotype; demonstrating that the transgene was able to rescue the embryonic lethal phenotype of the cyt cs KO. Lung fibroblasts (LF) from these animals were cultured and the transgenic cyt c cDNA deleted ex vivo by an adenovirus preparation expressing the Cre recombinase. clones by serial dilution and screened them for the presence of Cyt c via immunocytochemistry or western blot in which the resultant clone had no detectable Cyt c to generate the Cytochrome c Stable Knockout Mouse Fibroblast Cell Line.