C57BL/6-Rev3l^tm1Rsky/J

Stock number: 010540
Research areas: Research Tools

Description

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.

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.

Allele

Symbol: Rev3l^tm1Rsky
Name: targeted mutation 1, Klaus Rajewsky
MGI accession ID: MGI:3837724
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Rev3l
Marker name: REV3 like, DNA directed polymerase zeta catalytic subunit
Chromosome: 10
Strain of origin: B6.Cg-Thy1^a
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.