Cre4/2941#6 Cell Line
This cell line is a Lig3-/- derivative of the Cre4 cell line (Cat. ESA109). Cre4 cells were stably transfected with plasmid pMA2941, which encodes Cre recombinase. As a result of Cre-mediated excision, both Lig3 alleles in Cre4/2941#6 cells are inactive, which led to the loss of mtDNA.
From the laboratory of Mikhail F Alexeyev, PhD, University of South Alabama.
This cell line is a Lig3-/- derivative of the Cre4 cell line (Cat. ESA109). Cre4 cells were stably transfected with plasmid pMA2941, which encodes Cre recombinase. As a result of Cre-mediated excision, both Lig3 alleles in Cre4/2941#6 cells are inactive, which led to the loss of mtDNA.
From the laboratory of Mikhail F Alexeyev, PhD, University of South Alabama.
Specifications
| Product Type: | Cell Line |
| Cell Type: | Embryonic fibroblasts |
| Source: | Mouse E13.5 Embryo |
| Organism: | Mouse |
| Morphology: | Spindle-shaped at confluence |
| Biosafety Level: | BSL 1 |
| Subculturing: | Split 1:4 every 3 days |
| Growth Conditions: | DMEM/10% FBS/ 1 mM pyruvate, 50 µg/ml uridine, 1 mg/ml G418 |
| Cryopreservation: | DMEM/10% FBS/ 1 mM Pyruvate/50 µg/ml uridine /10% DMSO |
| Comments: | Slow- Growing |
| Storage: | LN2 |
| Shipped: | Dry Ice |
Provider
From the laboratory of Mikhail F Alexeyev, PhD, University of South Alabama.
Comments
Cre-TM mice (JAX 004682, B6.Cg-Tg(CAG-cre/Esr1)5Amc/J) were intercrossed with Lig3LoxP mice (Nature 471(2011) 240-244) to provide tamoxifen-inducible Cre expression driven from the actin promoter. Lig3Cre-TM mouse embryonic fibroblast (MEF) cell cultures were prepared from E13.5 embryonic mesenchyme. Cells were allowed to proliferate in T-25 tissue culture flasks at 37 °C in a humidified CO2-regulated (5%) incubator and were immortalized with a retroviral construct 3315, which encodes SV40 large T antigen (J. Biol. Chem. 288 (2013) 26594-26605) and selected depending upon their ability to form colonies. One clone was selected for its superior growth characteristics and designated Cre4. These cells were further transfected with a vector 2941, which encodes G418 resistance and CMV promoter driven, myc-tagged Cre recombinase to induce a deletion in the Lig3 gene. One of the resulting clones, Cre4/2941#6 underwent a biallelic loss of the Lig3, which was accompanied by the loss of mitochondrial DNA.
References
- Shokolenko IN, Fayzulin RZ, Katyal S, McKinnon PJ, Wilson GL, Alexeyev MF. Mitochondrial DNA ligase is dispensable for the viability of cultured cells but essential for mtDNA maintenance. J Biol Chem. 2013 Sep 13;288(37):26594-605.
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