B6J.Cg-Mettl5^em1.1Snac/J

Stock number: 037267
Product name: Mettl5 floxed

Research areas: Cell Biology Research, Neurobiology Research, Research Tools

Description

Exon 2 of the mouse Mettl5 (methyltransferase like 5) gene is flanked by loxP sites. Cre recombinase-mediated excision of the exon results in a knock-out allele useful in studies of methylation, mRNA translation, and metabolic dysregulation.

Of note, the donating investigator has also made a derived Mettl5 knock-out mouse line available (Stock No. 037268).

Detailed description

The METTL protein family is a class of S-adnosyl-methionine-dependent methyltransferases. Mutations in the human METTL5 (methyltransferase like 5) gene have been implicated in developmental abnormalities including microcephaly, intellectual disabilities, and attention deficit hyperactivity disorder (ADHD). METTL5 has also been shown to form a complex with TRMT112 to m⁶A methylate 18S rRNA in human cell lines and in mice, regulating mRNA translation.

Exon 2 of the mouse Mettl5 gene is flanked by loxP sites in this CRISPR/Cas9-generated allele. Cre recombinase-mediated excision of the floxed segment results in a knock-out allele.

Mice homozygous for a widespread knock-out allele (see Stock No. 037268) are sub-viable and fewer are born than predicted by Mendelian ratios. METTL5 expression is undetectable by qPCR, and greatly diminished by RNA-seq. Both male and female homozygous knock-out mice have a smaller body size and less body fat than their wildtype and heterozygous littermates throughout their lives.

Homozygotes do not demonstrate significant behavioral changes, as compared to heterozygous mice. RNA-seq analysis of brain and liver tissues reveals gene expression patterns that suggest dysregulated lipid, lipoprotein, and fatty acid metabolic pathways, as well as changes in thyroid hormone signaling.

Development

Exon 2 of the mouse Mettl5 gene was flanked by loxP sites using CRISPR/Cas9 methodologies in C57BL/6 x 129S6 F1-derived embryonic stem (ES) cells. An FRT-flanked neomycin cassette was excised. Resultant mice were backcrossed to C57BL/6J for 10 generations by the donating laboratory.

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: Mettl5^em1.1Snac
Name: endonuclease-mediated mutation 1.1, Sigrid Nachtergaele
MGI accession ID: MGI:7260126
Generation method: Endonuclease-mediated
Attributes: Conditional ready (e.g. floxed)
Marker symbol: Mettl5
Marker name: methyltransferase like 5
Chromosome: 2
Strain of origin: (C57BL/6 x 129S6/SvEvTac)F1
Molecular note: A Mettl5 conditional knockout line was made in which exon 2 was floxed, such that the presence of Cre would induce exon 2 deletion by creating a frame-shift mutation. An FRT-flanked neomycin cassette included in the construct was excised. The construct was electroporated with gRNA/Cas9 protein into ES cells that are F1 hybrid of C57bl/6 x 129S6.