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.

Catalog Number Product Size AVAILABILITY Price Qty
ESA110
Cre4/2941#6 Cell Line
1 vial In stock
Regular Price:$399.00
Specifications

Product Type: Cell Line
Cell Type: Embryonic fibroblasts
Organism: Mouse
Source: Mouse E13.5 Embryo
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
  1. 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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