Conditional miR-34afl/fl floxed mutant mice may have applications in studies related to the regulatory control of p53 pathways.
Andrea Ventura, Memorial Sloan Kettering Cancer Center
The miR-34 family of miRNAs (miR-34a, miR-34b, and miR-34c) are expressed mainly in the testis, brain, and lung. miR-34 proteins are induced upon p53 activation and are associated with tumor suppression, aging, neurodegeneration, and spermatogenesis. miR-34afl/fl mice possess loxP sites flanking the entire microRNA 34a (miR-34a) sequence. Homozygous miR-34afl/fl mice are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have miR-34a sequence deleted in the cre-expressing tissues. When crossed to mice expressing CAG-cre, miR-34a-/- mice exhibit abnormal neuronal precursor proliferation. When cre recombined miR-34a-/- mice are bred to Mirc21-/- mice (Stock No. 018546), which are deficient in miR-34b and miR-34c, the double mutant mice lack all three miR-34 family members. These miR-34 KO mice show normal development with an intact p53 pathway.
A targeting vector was designed to insert a loxP site upstream of the pre- microRNA 34a (miR-34a) sequence, and a frt-flanked neomycin resistance (neo) cassette, followed by a second loxP site, downstream of the pre-miR-34a sequence. The construct was electroporated into (C57BL/6 x 129S4/SvJae)F1-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and resulting chimeric mice were bred to Tg(ACTFLPe)9205Dym transgenic mice on a B6 background to delete the neo cassette. Progeny were crossed to remove the Flp-expressing transgene, resulting in a colony of miR-34afl/fl mice. These mice were maintained on mixed background. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
|Allele Name||targeted mutation 1.2, Andrea Ventura|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||targeted mutation 1.2, Andrea Ventura; Mir34atm1.2Aven|
|Gene Symbol and Name||Mir34a, microRNA 34a|
|Gene Synonym(s)||Mirn34a; mir 34a; Mirn34a; mmu-mir-34a|
|Strain of Origin||(C57BL/6 x 129S4/SvJae)F1|
|Molecular Note||A targeting vector was designed to insert a loxP site upstream of the sequence, and a frt-flanked neomycin resistance (neo) cassette, followed by a second loxP site, downstream of the sequence. Flp-mediated recombination removed the neo cassette and left the sequence floxed.|
|Mutations Made By|| |
Andrea Ventura, Memorial Sloan Kettering Cancer Center
When maintaining a live colony, homozygous mice may be bred together.
When using the miR-34afl mouse strain in a publication, please cite the originating article(s) and include JAX stock #018545 in your Materials and Methods section.
|Heterozygous or wildtype for Mir34a<tm1.2Aven>|
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