Overview

These catalytic subunit of DNA polymerase zeta (Rev3l) floxed mice may be useful in generating conditional mutations to study genome instability, DNA repair and class switch recombination.

Donating Investigator

Klaus Rajewsky, Max Delbruck Centre for Molecular Medicine

Genetic overview

Genetic Background	Generation

Rev3l^tm1Rsky

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), No functional change)	Rev3l	REV3 like, DNA directed polymerase zeta catalytic subunit

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Research Applications

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. When used in conjunction with a Cre recombinase-expressing strain, this strain is useful in generating tissue-specific mutants of the floxed allele. Excision of the floxed fragment in B cells results in chromosomal instability, impaired B cell proliferation, reduced class switch recombination, reduced frequency of somatic mutations and increased frequency of DNA breaks. This mutant mouse strain is useful in studies of genome instability, DNA break repair and class switch recombination.

For example, when crossed to a strain expressing Cre recombinase in Mature B cells (see Stock No. 006368), this mutant mouse strain may be useful in studies of class switch recombination and DNA break repair.

Development

A loxP flanked targeting vector containing neomycin resistance genes was utilized in the construction of this mutant. This selection cassette was inserted upstream of exon 2 of the targeted gene, and another loxP site was inserted downstream of exon 2. The construct was electroporated into C57BL/6 derived Bruce 4 embryonic stem (ES) cells. Transient Cre expression in targeted cells excised the neo cassette, but left the loxP-flanked exon 2. Correctly targeted ES cells were injected into blastocysts. The donating investigator reported that resulting chimeric animals were crossed to C57BL/6.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, at least 1 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Control Suggestions

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Schenten D; Kracker S; Esposito G; Franco S; Klein U; Murphy M; Alt FW; Rajewsky K. 2009. Pol zeta ablation in B cells impairs the germinal center reaction, class switch recombination, DNA break repair, and genome stability. J Exp Med 206(2):477-90PubMed: 19204108 MGI: J:146453

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Genetics

Rev3l^tm1Rsky

Allele Symbol: Rev3l^tm1Rsky

Allele Name	targeted mutation 1, Klaus Rajewsky
Allele Type	Targeted (Conditional ready (e.g. floxed), No functional change)
Allele Synonym(s)	Pol-zeta^f
Gene Symbol and Name	Rev3l, REV3 like, DNA directed polymerase zeta catalytic subunit
Gene Synonym(s)
Strain of Origin	B6.Cg-Thy1^a
Chromosome	10
Molecular Note	Exon 2 was targeted conditional deletion becuase messenger RNA splicing from exon 1 to exon 3 would result in a frameshift mutation. A floxed neomycin resistance cassette was placed upstream of the exon, and an additional loxP site was inserted downstream of the exon. After targeting, the neomycin cassette was removed by transient expression of Cre recombinase. Genomic PCR confirmed correct targeting of the locus.
Mutations Made By	Klaus Rajewsky, Max Delbruck Centre for Molecular Medicine

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Research Tools
- Cre-lox System
  - loxP-flanked Sequences

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Rev3l^tm1Rsky/Rev3l^tm1.1Rsky Tg(Cr2-cre)3Cgn/?
involves: C57BL/6

cellular phenotype

decreased B cell proliferation
- B cells stimulated in vitro in the presence of LPS or LPS + IL-4 have reduced cellular proliferation compared to controls
- viability of stimulated cells drops dramatically three days after stimulation

spontaneous chromosome breakage
- B cells undergoing class switch recombination have a 4-fold greater frequency of irregular chromosomes (broken chromosomes, chromosome fusions, and translocations) than controls
- many of these chromosome breaks occur at the IgH locus
- when B cells are activated without class switch recombination, there is still an increased frequency of chromosome breaks but none occur at the IgH locus

immune system phenotype

abnormal class switch recombination
- S-mu switch regions have less somatic mutations than controls due to mutant cells having less opportunity to accumulate mutations before switching
- there is a 2-3 fold reduced frequency of B cells that switch to IgG3 or IgG1 when stimulated in vitro in the presence of LPS or LPS + IL-4, respectively, as compared to controls
- this reduced class switch recombination is still observed when comparisons are made between B cells that have undergone the same number of divisions

abnormal somatic hypermutation frequency
- almost 50% of all sequences derived from these cells contained only one mutation, whereas close to 80% of the sequences derived from controls had multiple mutations
- the mutation percentage drops to less than a third of controls when germinal center B cells are pre-screened to keep only those that have excised the floxed allele
- the percentage of mutations that occur in the recombined IgH locus of germinal center B cells is roughly half that of controls

decreased B cell proliferation
- B cells stimulated in vitro in the presence of LPS or LPS + IL-4 have reduced cellular proliferation compared to controls
- viability of stimulated cells drops dramatically three days after stimulation

decreased germinal center B cell number
- there is a 2-3 fold reduction in the number of germinal center B cells
- this population is slightly enriched for cells that have escaped cre-mediated deletion of the floxed allele

decreased IgG1 level
- sera levels of antigen-specific IgG1 and Iglambda are half that of controls after immunization

hematopoietic system phenotype

abnormal class switch recombination
- S-mu switch regions have less somatic mutations than controls due to mutant cells having less opportunity to accumulate mutations before switching
- there is a 2-3 fold reduced frequency of B cells that switch to IgG3 or IgG1 when stimulated in vitro in the presence of LPS or LPS + IL-4, respectively, as compared to controls
- this reduced class switch recombination is still observed when comparisons are made between B cells that have undergone the same number of divisions

abnormal somatic hypermutation frequency
- almost 50% of all sequences derived from these cells contained only one mutation, whereas close to 80% of the sequences derived from controls had multiple mutations
- the mutation percentage drops to less than a third of controls when germinal center B cells are pre-screened to keep only those that have excised the floxed allele
- the percentage of mutations that occur in the recombined IgH locus of germinal center B cells is roughly half that of controls

decreased B cell proliferation
- B cells stimulated in vitro in the presence of LPS or LPS + IL-4 have reduced cellular proliferation compared to controls
- viability of stimulated cells drops dramatically three days after stimulation

decreased germinal center B cell number
- there is a 2-3 fold reduction in the number of germinal center B cells
- this population is slightly enriched for cells that have escaped cre-mediated deletion of the floxed allele

decreased IgG1 level
- sera levels of antigen-specific IgG1 and Iglambda are half that of controls after immunization

References

Schenten D; Kracker S; Esposito G; Franco S; Klein U; Murphy M; Alt FW; Rajewsky K. 2009. Pol zeta ablation in B cells impairs the germinal center reaction, class switch recombination, DNA break repair, and genome stability. J Exp Med 206(2):477-90PubMed: 19204108 MGI: J:146453

Additional - Rev3l^tm1Rsky related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Genotyping resources and troubleshooting
Breeding Considerations

While maintaining a live colony, these mice are bred as homozygotes.
- Additional Breeding and Husbandry Support
Citation
When using the C57BL/6-Rev3l^tm1Rsky/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #010540 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

Cryo Recovery

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Heterozygous for Rev3l<tm1Rsky>

Related Products and Services

Frozen Mouse Embryo	C57BL/6-Rev3l<tm1Rsky>/J	$2595.00
Frozen Mouse Embryo	C57BL/6-Rev3l<tm1Rsky>/J	$2595.00
Frozen Mouse Embryo	C57BL/6-Rev3l<tm1Rsky>/J	$3373.50
Frozen Mouse Embryo	C57BL/6-Rev3l<tm1Rsky>/J	$3373.50

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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

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Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Additional Information

Selected References

Rev3ltm1Rsky

Allele Symbol: Rev3ltm1Rsky

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Rev3ltm1Rsky/Rev3ltm1.1Rsky Tg(Cr2-cre)3Cgn/?involves: C57BL/6

cellular phenotype

immune system phenotype

hematopoietic system phenotype

References

Additional - Rev3ltm1Rsky related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Rev3l^tm1Rsky

Allele Symbol: Rev3l^tm1Rsky

Genotype: Rev3l^tm1Rsky/Rev3l^tm1.1Rsky Tg(Cr2-cre)3Cgn/?
involves: C57BL/6

Additional - Rev3l^tm1Rsky related