Prdm9EP knock in mice express a glutamic acid to proline amino acid (aa) substitution in mouse codon 360 of the Prdm9 (PR domain containing 9) locus on chromosome 17. The mutation creates a catalytic hypomorph; methyltransferase activity is severely reduced but not abolished. Male homozygotes are infertile and female homozygotes are subfertile. Prdm9EP mice may be useful for the study of meiotic recombination and fertility.
Kenneth Paigen, The Jackson Laboratory
Prdm9 (PR domain containing 9) encodes a meiosis-specific, DNA binding zinc finger protein that functions as a histone methyltransferase. Upon binding to DNA, PRMD9 catalyzes histone H3 lysines 4 and 36 trimethylation (H3K4me3 and H3K4me36) during meiotic prophase and is involved in controlling the positioning of recombination hotspots in humans and mice. Prdm9EP knockin mice were created using CRISPR/cas9 genome editing to express a glutamic acid to proline amino acid (aa) substitution in mouse codon 360 (initially reported as 365) of the Prdm9 locus on chromosome 17. The mutation creates a catalytic hypomorph; methyltransferase activity is severely reduced but not abolished. Prdm9 expression is not affected. Heterozygotes are viable and fertile. Homozygous males develop meiotic arrest and are infertile. Female homozygotes exhibit small ovaries, decreased numbers of oocytes and are subfertile. Most pachytene oocytes exhibit widespread asynapsis and unrepaired double stranded breaks. Prdm9EP mice may be useful for the study of meiotic recombination and fertility.
CRISPR/cas9 endonuclease-mediated genome editing was used to create an amino acid substitution at position 360 (glutamic acid to proline, Prdm9EP, GAG to CCG - initially reported as 365) in the PR/SET domain of the Prdm9 locus on chromosome 17. Single guide RNA and Cas9 endonuclease were introduced into C57BL/6J-derived fertilized eggs by pronuclear injection. The mutation was sequenced for confirmation. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to C57BL/6J mice for germline transmission. The colony was maintained on the C57BL/6J background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.
|Allele Name||endonuclease-mediated mutation 2, Kenneth Paigen|
|Allele Type||Endonuclease-mediated (Hypomorph)|
|Gene Symbol and Name||Prdm9, PR domain containing 9|
|Strain of Origin||C57BL/6J|
|Molecular Note||This catalytic hypomorph has a CRISPR/cas9-mediated GAG to CCG substitution resulting in a Glu360Pro (initially reported as 365) substitution in the PR/SET domain of C57BL/6J.|
When maintaining a live colony, heterozygous mice may be bred to each other or wildtype mice from the colony or C57BL/6J inbred mice (Stock No. 000664). Homozygous males are infertile and homozygous females are subfertile.
When using the Prdm9EP mouse strain in a publication, please cite the originating article(s) and include JAX stock #028854 in your Materials and Methods section.