B6.Cg-Mettl14^tm1.1Czhao/J

Stock number: 036375
Product name: Mettl14^f
Research areas: Cell Biology Research, Developmental Biology Research, Neurobiology Research, Research Tools

Description

Mettl14^f mutant mice possess loxP sites flanking exon 2 of the Mettl14 (methyltransferase like 14) gene and may be useful in generating conditional mutations to study histone modification and gene regulation particularly in embryonic neuronal stem cell development.

Detailed description

METTL14 (methyltransferase like 14) and METTL3 form a heterodimer that methylates RNA at the N⁶ position of adenosine. N⁶-methyladenosine or m⁶A is the most abundant internal mRNA modification that regulates diverse RNA activities. Within the heterodimer, METTL14 is required for substrate recognition and METTL3 conformation; METTL3 functions as the enzymatic subunit. These conditional Mettl14^f mice possess loxP sites flanking exon 2. Mice homozygous for the mutation are viable and fertile. Deletion of exon 2 creates an out-of-frame mutation that abolishes Mettl14 function.

Mettl14^f/f mice bred to mice with widespread Cre recombinase expression (B6.FVB-Tg(EIIa-cre)C5379Lmgd/J, Stock No. 003724) results in lethality of homozygous embryos during early embryogenesis.

Mettl14^f/f mice bred to mice with Cre recombinase expression in brain (B6.Cg-Tg(Nes-cre)1Kln/J, Stock No. 003771) results postnatal lethality by the first week of life. P0 brains exhibit an enlarged ventricle and a decrease in cortical thickness. Examination of embryonic brain reveals decreased proliferation and premature differentiation of cortical neural stem cells, and a decreased number of late-born neurons.

Development

The targeting construct was designed to contain from 5’ to 3’ – a loxP site, an FRT site, a splice acceptor, an IRES, hygromycin resistance cassette, a polyA sequence, a zeomycin resistance cassette, and an FRT site inserted into intron 1. A second loxP site is inserted in intron 2. Mettl14 is located on chromosome 3. The construct was electroporated into (129S6/SvEvTac x C57BL/6NCrl)F1-derived G4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. Chimeric males were crossed to C57BL/6 females and then crossed to B6;SJL-Tg(ACTFLPe)9205Dym/J mice to remove the hygro/zeo cassettes. The strain was backcrossed to C57BL/6J for 6 generations by the donating laboratory (see SNP note below). Upon arrival, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony.

In 2021, a 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the males sent to The Jackson Laboratory Repository. One of the 43 markers throughout the genome, on Chromosome 9, was segregating with SJL.

Allele

Symbol: Mettl14^tm1.1Czhao
Name: targeted mutation 1.1, Jing Crystal Zhao
MGI accession ID: MGI:6358166
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Synonyms: Mettl14^f
Marker symbol: Mettl14
Marker name: methyltransferase like 14
Marker synonyms: G430022H21Rik; hMETTL14; mKIAA1627; RGD1304822
Chromosome: 3
Strain of origin: (129S6/SvEvTac x C57BL/6NCrl)F1
Molecular note: The targeting vector introduced into intron 1 a loxP site, an FRT site, splice acceptor, IRES, hygromycin resistance cassette, polyadenylation sequence, zeomycin resistance cassette, and FRT site. An additional loxP site was isnerted downstream of exon 2. Flp-mediated recombination removed the selection cassettes and left exon 2 floxed.