These double knock-out mice have a neo cassettes replacing exons of the cyclin D binding myb-like transcription factor 1 (Dmtf1) and the cyclin-dependent kinase inhibitor 2A (Cdkn2d) genes. These mice may be useful for studying tumor suppression.
Kazushi Inoue, Wake Forest University School of Medicine
|Allele Type||Gene Symbol||Gene Name|
|Targeted (Null/Knockout)||Dmtf1||cyclin D binding myb-like transcription factor 1|
|Allele Type||Gene Symbol||Gene Name|
|Targeted (Modified isoform(s))||Cdkn2a||cyclin dependent kinase inhibitor 2A|
DMP1-/- ARF-/- double knock-out mice have a neo cassette replacing 2 exons of the cyclin D binding myb-like transcription factor 1 (Dmtf1) gene responsible for DNA-binding. They also have a neo cassette replacing exon 1β of the cyclin-dependent kinase inhibitor 2A (Cdkn2a or ARF) gene. DMP1 encodes a transcription factor activated by oncogenic Ras signaling that functions as a tumor suppressor through activation of the Arf-p53 pathway. CDKN2A encodes p19ARF, a cell cycle regulating protein that acts as a tumor suppressor via interaction with p53. Double homozygous mice may have an increased incidence of spontaneous tumors and lymphomas in the first year of life.
A targeting vector was designed to replace 2 exons of the cyclin D binding myb-like transcription factor 1 (Dmtf1) gene, involved in DNA-binding, with a neomycin resistance (neo) cassette. The construct was electroporated into 129X1/SvJ-derived RW-4 embryonic stem (ES) cells and correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting DMP1-/- chimeric males were bred to C57BL/6 females.
Another targeting vector was designed to replace exon 1β of the cyclin-dependent kinase inhibitor 2A (Cdkn2a) gene with a neo cassette. This construct was electroporated into 129X1/SvJ-derived RW-4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting ARF-/- chimeric males were bred to C57BL/6 females.
DMP1-/- mice and ARF-/- mice were bred together, and were subsequently backcrossed to C57BL/6 mice for at least 5 generations. Upon arrival at The Jackson Laboratory, double knock-out embryos were cryopreserved. These embryos were used to establish this live colony.
Some mice were also bred to C57BL/6J mice to separate the targeted mutations. The DMP1-/- single allelic strain is being maintained as Stock No. 029667. The ARF-/- single allelic strain is being maintained as Stock No. 029676).
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony of DMP1-/- ARF-/- mice at The Jackson Laboratory Repository. One of the 27 markers throughout the genome was segregating. Also, all 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the embryos sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.
|Allele Name||targeted mutation 1, Charles J Sherr|
|Allele Type||Targeted (Null/Knockout)|
|Allele Synonym(s)||targeted mutation 1, Charles J Sherr; Dmtf1tm1Cjs|
|Gene Symbol and Name||Dmtf1, cyclin D binding myb-like transcription factor 1|
|Gene Synonym(s)||DMP1; DMTF; hDMP1; MRUL; Dmp1|
|Strain of Origin||129X1/SvJ|
|Molecular Note||Exons encoding residues involved in DNA-binding were disrupted by the insertion of a 1.0kb neomycin selection cassette. Western blot analysis using antibodies raised against the carboxy terminal and the Myb-repeat domain showed an absence of encoded protein in homozygous mutant mice.|
|Allele Name||targeted mutation 1, Charles J Scherr|
|Allele Type||Targeted (Modified isoform(s))|
|Allele Synonym(s)||Cdkn2atm1Cjs; targeted mutation 1, Charles J Scherr|
|Gene Symbol and Name||Cdkn2a, cyclin dependent kinase inhibitor 2A|
|Gene Synonym(s)||ARF; CMM2; ARF-INK4a; MTS-1; MTS1; CDKN2; MLM; INK4; INK4A; INK4a-ARF; Ink4a/Arf; P14; P14ARF; P16; P16INK4; p16Cdkn2a; P19; p19ARF; P19ARF; Pctr1; P16-INK4A; P16INK4A; Pctr1; CDK4I; TP16; plasmacytoma resistance 1; Arf; p16INK4a; p19ARF; p16|
|Strain of Origin||129X1/SvJ|
|General Note||Phenotypic Similarity to Human Syndrome: Rhabdomyosarcoma, Embryonal J: 237183 in mice homozygous for Cdkn2atm1Cjs and hemizygous for Tg(CKMM-tTA)A3Rhvh and Tg(tetO-Hgf,-EGFP)24Tcre.|
|Molecular Note||Replacement of exon 1 beta of the Cdkn2a gene, encoding the translation start site for alternative splice transcript p19ARF, with a neomycin cassette. Note that expression of alternative splice transcript p16Ink4a, transcribed from exon 1 alpha, is not affected by this mutation.|
When maintaining a live colony, double homozygous mice may be bred together.
When using the DMP1-/- ARF-/- mouse strain in a publication, please cite the originating article(s) and include JAX stock #029709 in your Materials and Methods section.
|Heterozygous or wildtype for Dmtf1<tm1Cjs> and Cdkn2a<tm1Cjs>|
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