MB19tsA, clone 2B2 [Mbeta19tsA, clone 2B2] (ATCC® CRL-2308)

Organism: Mus musculus, mouse  /  Cell Type: fibroblastSV40 large T antigen transfected  / 

Permits and Restrictions

View Permits

Organism Mus musculus, mouse
Cell Type fibroblastSV40 large T antigen transfected
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 SV-40 virus (Cells contain SV-40 viral sequences)
Age 10.5 days gestation
Applications
MB19tsA, clone 2B2 is a fibroblast-like cell line derived from primary cultures of DNA polymerase beta (pol-beta) deficient fibroblasts isolated from embryos from heterozygous females ( beta-polymerase +/-) at day 10.5 of gestation.
The cell line is deficient in base excision repair due to homozygous deletion of the promoter and exon I in the pol-bet a gene.
This deletion event is reversible and therefore this cell line is ideal for site-specific recombination using the Cre-lox system.
The contributor of this line has demonstrated that insertion of a single-copy of a variety of cDNAs can be site-specifically reinserted at the pol-beta locus, and that this is a stable event.
Derivation
MB19tsA, clone 2B2 is a fibroblast-like cell line derived from primary cultures of DNA polymerase beta (pol-beta) deficient fibroblasts isolated from embryos from heterozygous females ( beta-polymerase +/-) at day 10.5 of gestation.
Clinical Data
MB19tsA, clone 2B2 is a fibroblast-like cell line derived from primary cultures of DNA polymerase beta (pol-beta) deficient fibroblasts isolated from embryos from heterozygous females ( beta-polymerase +/-) at day 10.5 of gestation.
Comments
The genotype of this cell line has recently been determined to be DNA polymerase beta mutant (null) and DNA polymerase iota mutant (null).
MB19tsA, clone 2B2 is a fibroblast-like cell line derived from primary cultures of DNA polymerase beta (pol-beta) deficient fibroblasts isolated from embryos from heterozygous females ( beta-polymerase +/-) at day 10.5 of gestation.
The pol-beta deficient fibroblasts were immortalized by transfection with the DNA plasmid construct ptsA58H which contains coding sequences for both a hygromycin-resistance gene and tsA58H, a temperature-sensitive SV40 T antigen.
Cells were selected in the presence of 0.080 mg/ml hygromycin for 21 days and further selected by limiting dilution.
The cell line is deficient in base excision repair due to homozygous deletion of the promoter and exon I in the pol-bet a gene.
The deletion of pol-beta is mediated by the Cre-loxP recombination.
This deletion event is reversible and therefore this cell line is ideal for site-specific recombination using the Cre-lox system.
The contributor of this line has demonstrated that insertion of a single-copy of a variety of cDNAs can be site-specifically reinserted at the pol-beta locus, and that this is a stable event.
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and supplemented with 0.08 mg/ml hygromycin B, 90%; fetal bovine serum, 10%
Subculturing
Subcultivation Ratio: A subcultivation ratio of 1:10 to 1:15 is recommended
Medium Renewal: Every 2 to 3 days
Remove medium, rinse quickly with 0.25% trypsin, 0.03% EDTA solution, add 1-2 ml additional trypsin solution and allow flasks to set at room temperature (or at 37C) until the cells detach. Add fresh culture medium, aspirate and dispense into new flasks.
Cryopreservation
Culture medium, 95%; DMSO, 5%
Culture Conditions
Temperature: 34.0°C
Name of Depositor RW Sobol, SH Wilson
References

Gu H, et al. Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. Science 265: 103-106, 1994. PubMed: 8016642

Sobol RW, et al. Requirement of mammalian DNA polymerase-beta in base-excision repair. Nature 379: 183-186, 1996. PubMed: 8538772

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Gu H, et al. Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. Science 265: 103-106, 1994. PubMed: 8016642

Sobol RW, et al. Requirement of mammalian DNA polymerase-beta in base-excision repair. Nature 379: 183-186, 1996. PubMed: 8538772