These mitogen-activated protein kinase 14 (Mapk14) knock-in (or p38AF) mice harbor dominant-negative mutations of the activating phosphorylation sites of the p38MAPK (now Mapk14) gene that result in specifically attenuated p38MAPK signaling. These p38AF mice may be useful in studying cell cycle inhibitors, cellular senescence, ageing, and cell proliferation/regeneration in response to stress.
Dmitry V Bulavin, Institute for Research on Cancer and Aging, Nice (IRCAN)
Mice heterozygous for the p38AF dominant-negative allele are viable and fertile, with two amino acid substitutions (T180A and Y182F) at activating phosphorylation sites of the p38MAPK locus. Homozygous embryos die around embryonic day 11.5 with placental and heart defects (similar to some p38 knockout mice). Mice heterozygous for the p38AF dominant-negative allele exhibit specifically attenuated p38MAPK signaling without any reported effects on other MAPK pathways. Heterozygous mice do not exhibit any accelerated tumorigenesis on this genetic background (in fact, heterozygous mice crossed into two different tumor-prone genetic backgrounds also show no increased tumorigenic potential). Heterozygous mice show reduced aging-dependent activation of multiple cell cycle inhibitors in multiple tissues without increasing cancer predisposition. Specifically, reduced inhibitors include the products of the Cdkn2a tumor suppressor locus (p16Ink4a and p19Arf), as well as p15Ink4b and p21Waf1. Aged heterozygous mice have improved proliferation and regeneration in pancreatic islets (compared to wildtype). Recovery from streptozocin induced pancreatic beta cell depletion is significantly improved in heterozygotes (compared to similarly treated wildtype mice).
To generate the p38AF dominant-negative mutation, a mouse p38MAPK (p38&alpha or Mapk14) DNA sequence was modified via site-directed mutagenesis to harbor two amino acid substitutions at activating phosphorylation sites, threonine 180 to alanine (T180A) and tyrosine 182 to phenylalanine (Y182F). A targeting vector with these polymorphisms in exon 7, a new BamH1-specific restriction site in the preceding intron, and a loxP-flanked PGK-neo cassette downstream of exon 8 was electroporated into C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred with C57BL/6 females to establish the colony. Heterozygous mice were then bred with EIIa-cre mice (on a C57BL/6 congenic background). The resulting mice with the floxed neo cassette removed were then selectively bred together to remove the EIIa-cre transgene. Mice heterozygous for the p38AF allele were bred together or to wildtype siblings or to C57BL/6J inbred mice for many generations prior to sending to The Jackson Laboratory Repository. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
|Expressed Gene||Mapk14, mitogen-activated protein kinase 14, mouse, laboratory|
|Site of Expression|
|Allele Name||targeted mutation 1.1, Dmitry V Bulavin|
|Allele Synonym(s)||p38AF; p38alphaki|
|Gene Symbol and Name||Mapk14, mitogen-activated protein kinase 14|
|Promoter||Mapk14, mitogen-activated protein kinase 14, mouse, laboratory|
|Expressed Gene||Mapk14, mitogen-activated protein kinase 14, mouse, laboratory|
|Strain of Origin||C57BL/6|
|Molecular Note||Regulatory sites Thr180 and Tyr182 are replaced by alanine and phenylalanine, respectively. A floxed neo cassette inserted downstream of exon 8 was removed by germ line, cre mediated recombination.|
|Mutations Made By|| |
Dmitry Bulavin, Institute for Research on Cancer and Aging, Nice (IRCAN)
When maintaining a live colony, heterozygous mice may be bred to wildtype mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). Homozygous embryos die around embryonic day 11.5 with placental and heart defects.
When using the B6.Cg-Mapk14tm1.1Dvb/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #012736 in your Materials and Methods section.
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