Prmt6-/- KO mice lack the entire coding region of the protein arginine N-methyltransferase 6 (Prmt6) gene making them useful for studying the role of arginine methylation during cellular senescence.
Stephane Richard, McGill University
Prmt6-/- KO mice lack the entire coding region of the protein arginine N-methyltransferase 6 (Prmt6) gene, abolishing gene expression. Mice homozygous for this allele are viable and fertile. PRMT6 mediates the asymmetric methylation of arginine 2 of histone H3 ally [H3R2(me2a)]. It acts as a transcriptional repressor of various genes such as Trp53 (transformation related protein 53), and also as a transcriptional coactivator of a number of steroid hormone receptors. Cultured mouse embryonic fibroblasts (MEFs) from these mice exhibit cell cycle arrest at G0/G1, with morphological changes such as cell flattening and enlarged cytoplasm evident. Prmt6-/- MEFs show an upregulation of the ARF-p53 pathway and an upregulation of cyclin-dependent kinase (CDK) inhibitors such as p21.
A targeting vector was designed by Dr. Stephane Richard (McGill University) to insert loxP sites flanking the protein arginine N-methyltransferase 6 (Prmt6) coding region. A frt-flanked neomycin resistance (neo) cassette, in opposite orientation to the gene, was placed downstream of the floxed region. The construct was electroporated into (129X1/SvJ x 129S1/Sv)F1-Kitl+
-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts and resulting chimeric mice were crossed to B6.129S4-Gt(ROSA)26Sortm2(FLP*)Sor/J mice (Stock No. 012930) to remove the neo cassette. Mice were subsequently bred to B6.FVB-Tg(EIIa-cre)C5379Lmgd/J mice (Stock No. 003724) to remove the Prmt6 coding region. Resulting mice were crossed to remove the Flp- and Cre-expressing transgenes. Prmt6-/- mice were bred to C57BL/6NCrl mice for at least 12 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).
|Allele Name||targeted mutation 1.2, Stephane Richard|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Prmt6, protein arginine N-methyltransferase 6|
|Strain of Origin||129|
|Molecular Note||A loxP site was inserted upstream of exon 1. A loxP- and FRT-flanked neomycin resistance cassette was inserted downstream of exon 1. Flp-mediated recombination removed the neomycin resistance cassette and left exon 1 floxed. Cre-mediated recombination removed exon 1. Western blot analysis confirmed the absence of protein expression in mouse embryonic fibroblasts.|
When maintaining a live colony, homozygotes may be bred together.
When using the Prmt6- mouse strain in a publication, please cite the originating article(s) and include JAX stock #028929 in your Materials and Methods section.