This murine fibroblast cell line is a derivative of the 4B6 cell line (Cat. ESA102). Unlike the parental cell line, it expresses the Escherichia coli Lig A gene in place of the endogenous DNA ligase III. To generate 4B6/3598/3442#2 cells, 4B6 cell line was transduced with retroviruses rv3598 (G418 resistance) and rv3442 (puromycin resistance) resulting in the inactivation of the cellular Lig 3 gene through Cre-mediated excision (rv3442) and re-expression of the bacterial LigA gene devoid of mitochondrial targeting signal (rv3598). Inefficient mitochondrial import of the LigA in the absence of the Lig 3 makes DNA ligase activity limiting for the mtDNA replication, and leads to a reduced (10-100 fold) mtDNA copy number.
From the laboratory of Mikhail F Alexeyev, PhD, University of South Alabama.
|Product Type:||Cell Line|
|Cell Type:||Embryonic fibroblasts|
|Morphology:||Spindle-shaped at confluence|
|Source:||Mouse E13.5 Embryo|
|Biosafety Level:||BSL 1|
|Growth Conditions:||DMEM/10% FBS. Periodic selection with 10 µg/ml blasticidin, 2 µg/ml puromycin, and 1 mg/ml G418|
|Subculturing:||Split 1:4 every 3 days|
|Cryopreservation:||DMEM/10% FBS/10% DMSO|
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