These floxed mutant mice possess loxP sites flanking exons 2, 3 and 4 of the Tsc2 gene. This strain may be useful for generating conditional mutations in applications related to the study of tuberous sclerosis complex.
Michael Gambello, Emory University
The Tsc2 gene encodes the tuberin protein which interacts with hamartin. Mutations in the TSC2 gene can cause tuberous sclerosis complex.
These Tsc2flox mice possess loxP sites flanking exons 2, 3 and 4 of the targeted gene. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exons 2, 3 and 4 deleted in the cre-expressing tissues.
When bred to a strain with widespread Cre recombinase expression (transgenic CMV-cre mice), the resulting recombined allele has exons 2, 3 and 4 completely deleted.
A targeting vector containing a loxP-frt-neo-frt cassette was utilized in the construction of this mutant. This selection cassette was inserted into intron 1 (upstream of exon 2) of the targeted gene, and another loxP site was inserted downstream of exon 4. This construct was electroporated into 129-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were crossed to albino C57BL/6. Heterozygote Tsc2+/neo mice were crossed with transgenic mice expressing Flpe recombinase under the control of the beta actin promoter (Tg(ACTFLPe)9205Dym) to remove the FRT flanked NEO cassette. Mice that retained the loxP site flanked exons 2, 3 and 4 were then bred to remove the Tg(ACTFLPe)9205Dym transgene. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
|Allele Name||targeted mutation 1.1, Michael J Gambello|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Tsc2, tuberous sclerosis 2|
|Strain of Origin||(129X1/SvJ x 129S1/Sv)F1-Kitl+|
|Molecular Note||This allele contains loxP sites flanking exons 2, 3 and 4. A frt-flanked neomycin selection cassette was removed from the first intron by crossing with mice expressing FLP recombinase in the germline, leaving a single frt site downstream of the first loxP site.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the Tsc2flox mouse strain in a publication, please cite the originating article(s) and include JAX stock #027458 in your Materials and Methods section.