B6;SJL-Plk4^em2Ahol/J

Stock number: 037549
Product name: Plk4^f
Strain synonyms: Plk4^flox
Research areas: Cell Biology Research, Research Tools

Description

Plk4 ^f mice possess loxP sites flanking exon 5 of the Plk4 (Polo-like kinase 4) gene. These mice may be useful for studying the role of Plk4 in centriole biology and centrosome loss in multiple disorders.

Detailed description

Polo-like kinase 4 (PLK4) is a serine/threonine kinase that regulates duplication of centrioles during cell-cycle progression. Overexpression of PLK4 may result in centriole amplification which is a common feature of cancer cells and other chromosomal instability disorders, while PLK4 loss causes centrosome duplication failure. Mice that are homozygous for this floxed allele are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 5 deleted in the cre-expressing tissues. These Plk4 ^f mice may be useful for studying the role of Plk4 in centriole biology and centriole loss in specific tissues.

The donating investigator has made multiple Plk4 mouse lines available: Plk4^Δu (Stock No. 037482) Plk4^Dox (Stock No. 037530) and Plk4^f (Stock No. 037549).

Development

The Plk4^f allele was generated using CRISPR/cas9 endonuclease-mediated homology-directed repair (HDR) genome editing. Two guide RNAs (mPLK4 sgRNA1: 5'-aagctaggactttaatactc-3', mPLK4 sgRNA2: 5'-ctgcatgtagagggaagctg-3') were used to target the introns flanking exon 5 of the Plk4 gene. Plk4 transcript Plk4-201 was used as reference for the exon number, and the guide and donor sequences. The two sgRNAs, ssDNA (repair template introduced two loxP sites into intron 4 and 5), and the Cas9 nuclease mRNA were introduced into the cytoplasm of zygotes derived from B6SJLF1 mice with well recognized pronuclei. Correctly targeted pups were identified by PCR and sequencing. The resulting Plk4^f/+ mice were backcrossed to C57BL/6J mice for one generation to minimize off-target CRISPR effects by the donating laboratory. The donating investigator reports their colony exhibits white/albino, agouti and black coat colors (as expected for a mixed C57BL/6;SJL genetic background). Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Plk4^em2Ahol
Name: endonuclease-mediated mutation 2, Andrew Holland
MGI accession ID: MGI:7316651
Generation method: Endonuclease-mediated
Attributes: Conditional ready (e.g. floxed)
Synonyms: Plk4^f
Marker symbol: Plk4
Marker name: polo like kinase 4
Marker synonyms: MCCRP2; Sak; Stk18
Chromosome: 3
Strain of origin: (C57BL/6J x SJL/J)F1
Molecular note: CRISPR/cas9 genome editing uses two sgRNAs RNAs (mPLK4 sgRNA1: 5'-aagctaggactttaatactc-3', mPLK4 sgRNA2: 5'-ctgcatgtagagggaagctg-3') and ssDNA to introduce two loxP sites into intron 4 and 5.