B6;129S-Trp53^tm6.1Glo/J

Stock number: 039758
Product name: Trp53^wm-R245W
Research areas: Apoptosis Research, Cancer Research, Research Tools

Description

Trp53^wm-R245W mice allow Cre recombinase-inducible expression of Trp53*R245W, a transformation related protein 53 (Trp53) with a point mutation in codon 245 (CGC->TGG; Arg245Trp), corresponding to the human TP53*R248W missense mutation. Following Cre exposure, these mice may be useful for tracing mutant p53-depedent cell proliferation, modified cell membranes, increased cell invasion, enhanced cell survival, and metastasis in cancer.

Of note, the donating investigator has also made available their Trp53^wm-R172H mice (Stock No. 039672).

Detailed description

The Trp53^wm-R245W allele normally expresses wild-type p53, but, upon Cre-mediated recombination, expresses the missense mutation form of Trp53R245W, transformation related protein 53 (Trp53) with a point mutation in codon 245 (CGC->TGG; Arg245Trp; wm refers to wild type to mutant). When crossed with cell/tissue-specific Cre-expressing mice, the model mimics somatic p53 mutagenesis event by inducing conditional mutant Trp53R245W in adult cells/tissues. The heterozygous and homozygous mice are viable and fertile without Cre-recombination. There are no exceptional features before Cre-recombination. The Trp53^wm-R245W allele expresses a full-length p53 protein and maintains normal function in heterozygous and homozygous mice.

Of note, in addition to these mice, The Jackson Laboratory Repository has Trp53^wm-R172H mice (Stock No. 039672).

Development

The targeting vectors were generated by cloning a cDNA fragment that contained exons 7-11 of the wildtype Trp53 and polyadenylation signal, followed by an Frt-flanked PGKneo cassette into intron 6 of a Trp53 genomic fragment. LoxP sequences flanked the cDNA fragment and PGKneo cassette. An arginine to tryptophan (C-to-T change at position 733 and C-to-G change at position 735) substitution at codon 245 (p.R245W) in exon 7 was performed by site-directed mutagenesis. The resulting constructs were then individually cloned into a vector containing a thymidine kinase (TK) cassette. The entire targeting vectors were sequence verified, linearized, and electroporated into 129S7/SvEvBrd-Hprt⁺ derived AB-1 embryonic stem (ES) cells. Two targeted ES cell clones were injected into recipient blastocysts and transferred into pseudo-pregnant female mice. The resulting chimeras were mated with C57BL/6J females to secure germline transmission of the mutant alleles. Mice with the targeted alleles were crossed with Rosa26-flp mice (Stock No. 009086) to remove the PGKneo cassettes. The resulting mice were backcrossed to C57BL/6J mice at least two generations and maintained on a mixed C57BL/6;129S strain background prior to sending to The Jackson Laboratory in April 2025. Upon arrival, the mice were bred at least one generation to C57BL/6J (Stock No. 000664), and then subsequently cryopreserved as heterozygous sperm in September 2025.

Allele

Symbol: Trp53^tm6.1Glo
Name: targeted mutation 6.1, Guillermina Lozano
MGI accession ID: MGI:7341396
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Synonyms: Trp53^wm-R245W
Marker symbol: Trp53
Marker name: transformation related protein 53
Marker synonyms: BCC7; BMFS5; LFS1; p44; p53; TRP53
Chromosome: 11
Strain of origin: 129S7/SvEvBrd-Hprt1⁺
Molecular note: An arginine to tryptophan (C-to-T change at position 733 and C-to-G change at position 735) substitution was introduced at residue 245 (p.R245W) in exon 7. In addition, a loxP site, Trp53 exons 7-11 with a native polyadenylation signal sequence, and FRT site flanked PGK neomycin resistance gene cassette and second loxP site were inserted in intron 6. The PGK neomycin cassette was removed via Flp-mediated recombination.