B6N.129S-Kat5^tm1.1Ge/J

Stock number: 035014
Product name: TIP60^flox

Research areas: Neurobiology Research, Research Tools

Description

TIP60 floxed mice have loxP sites flanking exons 1-9 of the K(lysine) acetyltransferase 5 (Kat5) gene. Exposure to Cre recombinase removes the floxed sequence - creating a null allele. These TIP60 floxed mice may be useful in the study of circadian rhythms and related diseases.

Detailed description

The Kat5 gene (also known as Tip60) encodes a lysine acetyltransferase which acetylates the BRD4 binding partner BMAL1 and plays a critical role in the regulation of circadian rhythm.

The TIP60 floxed allele has loxP sites flanking exons 1-9 of the Kat5 gene. Prior to introduction of Cre recombinase, mice heterozygous or homozygous for the TIP60 floxed allele are viable and fertile with normal breeding, and have no gross physical or behavioral abnormalities.

When TIP60 floxed are bred to mice that express Cre recombinase, the resulting offspring will have exons 1-9 deleted - creating a null allele in the cells/tissues as determined by the Cre promoter.

For example, when bred to a neural-specific Cre (Syt10-cre), mice homozygous for the deletion of TIP60 in cells of the suprachiasmatic nucleus exhibit reduced expression of clock genes and altered locomotor behavior in a constant darkness environment (Petkau et al. 2019).

Development

The TIP60 floxed allele (also called Kat5^FL) was created in the laboratory of Dr. Gregor Eichele (Max Planck Institute for Biophysical Chemistry). First, A 15.7 kb fragment of Tip60 genomic DNA was cloned from a BAC clone (bMQ-331N14, Sanger Institute) into a targeting vector carrying a Pol2-DTA cassette for negative selection. The first loxP site was inserted 637 bp upstream of the Tip60 ATG start codon. The second loxP site together with a FRT-flanked PGK-neo cassette was inserted 3,065 bp downstream of the ATG. The 5' end of the homologous arm of the targeting vector was 4.5 kb and the 3' end of the homologous arm was 7.6 kb. The construct was electroporated into 129S6/SvEvTac-derived TC-1 ES cells. Correctly targeted ES cells were then microinjected into recipient blastocysts and chimeric mice were crossed to C57BL/6NCrl for germline transmission. The FRT-flanked PGK-neo selection cassette was then removed by crossing with Flpe-expressing mice (Stock No. 003946). The strain was subsequently backcrossed to C57BL/6N for at least 20 generations prior to sending males to The Jackson Laboratory in 2020. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

Allele

Symbol: Kat5^tm1.1Ge
Name: targeted mutation 1.1, Gregor Eichele
MGI accession ID: MGI:6356678
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: Tip60^flox
Marker symbol: Kat5
Marker name: K(lysine) acetyltransferase 5
Marker synonyms: cPLA2; ESA1; HTATIP; HTATIP1; mHTATIP/CPLA2; NEDFASB; PLIP; TIP; TIP60; ZC2HC5
Chromosome: 19
Strain of origin: 129S/SvEv
Molecular note: Exons 1-9 were flanked by loxP sites and an frt-flanked neomycin selection cassette. The neomycin selection cassette was removed via FLP-mediated recombination in the germline.