4B6/3598/3442#2 Cell Line
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.
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 |
| Organism: | Mouse |
| 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 |
| Comments: | Slow- Growing |
| Storage: | LN2 |
| Shipped: | Dry Ice |
- Spadafora D, Kozhukhar N, Alexeyev MF. Presequence-Independent Mitochondrial Import of DNA Ligase Facilitates Establishment of Cell Lines with Reduced mtDNA Copy Number. PLoS One. 2016 Mar 31;11(3):e0152705.
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