ARF-/- mice have a neo cassette replacing exon 1β of the cyclin-dependent kinase inhibitor 2A (Cdkn2a) gene. These mice may be useful for studying tumor suppression.
Kazushi Inoue, Wake Forest University School of Medicine
ARF-/- mice have a neo cassette replacing exon 1β of the cyclin-dependent kinase inhibitor 2A (Cdkn2a) gene, abolishing gene function. CDKN2A encodes p19ARF, a cell cycle regulating protein that acts as a tumor suppressor via interaction with p53. ARF-/- mice have an increased incidence of tumors in the first year of life, with an increase seen in the second year. Mean latency of spontaneous tumors in homozygous knock-out mice is 38 weeks. Sarcomas, lymphomas, carcinomas, central nervous system tumors are common.
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 a live colony. The DMP1-/- allele was bred out of this colony of ARF-/- mice, and is being maintained as a separate strain (Stock No. 029667).
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 Scherr|
|Allele Type||Targeted (Modified isoform(s))|
|Allele Synonym(s)||ARF-; p19ARF-; p19Arf-null; p19Arf-|
|Gene Symbol and Name||Cdkn2a, cyclin dependent kinase inhibitor 2A|
|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, homozygous mice may be bred together.
When using the Arf KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #029676 in your Materials and Methods section.
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