STOCK Cdkn2a^tm3(dre)Bzsh/J

Stock number: 039749
Product name: p16-LSL-Dre
Research areas: Cancer Research, Research Tools

Description

The p16-LSL-Dre knock-in mice express Dre recombinase from the endogenous cyclin dependent kinase inhibitor 2A (Cdkn2a) gene also known as p16INK4a after exposure to Cre recombinase. A floxed-STOP cassette prevents Dre expression without Cre recombinase activity. These mice are useful for intersectional genetic systems to study the roles of cellular senescence in physiological and pathological processes, including embryonic development, cancer, aging, and multiple other diseases.

Detailed description

The p16-LSL-Dre knock-in mice have a loxP-stop-loxP (or LSL) in front of the P2A-Dre cassette inserted between the last exon and 3'UTR of the cyclin dependent kinase inhibitor 2A (Cdkn2a) gene, also known as p16INK4a. Insertion of this LSL-Dre cassette does not affect endogenous Cdkn2a gene expression. Cdkn2a encodes a cyclin-dependent kinase (CDK) inhibitor protein expressed in senescent cell populations. The expression of p16 INK4a is essentially undetectable in young and healthy tissues, but p16 INK4a+ cells accumulate in many aged or diseased tissues. When crossed to cell/tissue-specific Cre-recombinase expressing mice, intersectional genetic tracing can be achieved in these mice. Homozygous mice are viable and fertile.

Development

A targeting vector containing the triple SV40 polyadenylation (tpA) that functions as transcription terminators (stop) flanked by two loxP sites (loxP-stop-loxP or LSL), self-cleaving peptide 2A sequence, and a complementary DNA (cDNA) encoding Dre recombinase followed by a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), polyA signal, and FRT-flanked neomycin resistance cassette was inserted into the last exon of the Cdkn2a gene replacing the stop codon. The construct was electroporated into unspecified embryonic stem (ES) cells derived from the C57BL/6x129S3 genetic background. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were crossed to C57BL/6J, and offspring were crossed to ICR to maintain under the C57BL/6;ICR mixed background by the donating laboratory. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Cdkn2a^tm3(dre)Bzsh
Name: targeted mutation 3, Bin Zhou
MGI accession ID: MGI:8172891
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Recombinase-expressing
Marker symbol: Cdkn2a
Marker name: cyclin dependent kinase inhibitor 2A
Marker synonyms: Arf; ARF-INK4a; CAI2; CDK4I; CDKN2; CMM2; INK4; INK4A; Ink4a/Arf; INK4a-ARF; MLM; MTS1; MTS-1; P14; P14ARF; P16; p16Cdkn2a; P16INK4; P16INK4A; P19; p19^ARF; P19ARF; Pctr1; TP16
Chromosome: 4
Strain of origin: (C57BL/6 x 129S)F1
Molecular note: A targeting vector containing the triple SV40 polyadenylation (tpA) that functions as transcription terminators (stop) flanked by two loxP sites (loxP-stop-loxP or LSL), self-cleaving peptide 2A sequence and a complementary DNA (cDNA) encoding Dre recombinase followed by a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), polyA signal, and FRT-flanked neomycin resistance cassette was inserted into the last exon of the Cdkn2a gene replacing the stop codon.