STOCK Mettl3^tm1.1Jcbr/MgkJ

Stock number: 040326
Product name: Mettl3 f/f
Strain synonyms: Mettl3 flox/flox
Research areas: Cancer Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Mettl3 f/f conditional knock-out mice have exon 4 of the methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit (Mettl3) gene flanked by loxP sites. When crossed with tissue-specific Cre mice, the resulting homozygous offspring will have Mettl3 knock-out in Cre-expressing tissues. These mice may be useful in studies of METTL3 and RNA methylation.

Detailed description

Mettl3 (methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit) catalyzes the post-transcriptional mRNA methylation to N6-methyladenosine (m6A). The presence of m6A alters mRNA metabolism, including mRNA stability, mRNA splicing, and translation efficiency. Mettl3 f/f conditional knock-out mice have Mettl3 exon 4 flanked by loxP sites. Heterozygous and homozygous mice are viable and fertile with a healthy, normal appearance. When bred to mice expressing tissue-specific Cre recombinase, offspring will have exon 4 deleted in the Cre-expressing tissues. These Mettl3 flox mice may be useful for studying the role of Mettl3 in temporal/spatial-specific mRNA methylation.

Development

The targeting vector obtained from the KOMP Repository at UC Davis by Dr. Jens Brüning (Max Planck Institute for Metabolism Research) was used to make Knock-Out first allele, which is essentially identical to Mettl3^{tm1a(KOMP)Wtsi}. The targeting construct incorporates an FRT-lacZ-loxP followed by a neomycin resistance gene, SV40 polyA, a second FRT site, and a second loxP site upstream of exon 4 at a position 52,535,930 of Chromosome 14 of the Mettl3 gene, and a third loxP site inserted downstream in intron 4 at position 52,536,652 (Build GRCm39). The construct was electroporated into unspecified embryonic stem (ES) cells. Targeted ES cells were injected into blastocysts to generate chimeric mice. Those mice are then crossed to unspecified germline Flp mice to remove the lacZ sequence and neo selection cassette, leaving loxP sites flanking the exon 4 to generate a "conditional ready" (floxed) allele. Subsequently, Dr. Michael Kharas (Memorial Sloan Kettering Cancer Center) maintained the Mettl3 f/f mice that had the Flp allele bred out. Dr. Kharas provides those mice to The Jackson Laboratory. Upon arrival, sperm is cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Mettl3^tm1.1Jcbr
Name: targeted mutation 1.1, Jens C Bruning
MGI accession ID: MGI:7856631
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: Mettl3 f; Mettl3 flox
Marker symbol: Mettl3
Marker name: methyltransferase like 3
Marker synonyms: 2310024F18Rik; hMETTL3; IME4; M6A; MT-A70; Spo8
Chromosome: 14
Strain of origin: Not Specified
Molecular note: A "conditional ready" (floxed) allele was created by flp recombinase expression in mice carrying Mettl3^tm1Jcbr allele to remove the lacZ sequence and neo selection cassette, leaving loxP sites flanking the critical exon(s). The L1L2_Bact_P cassette was inserted at position 52535930 of Chromosome 14 upstream of the critical exon(s) (Build GRCm39). The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by a neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of the targeted exon 4 at position 52536652. After flp recombination, the critical exon 4 is thus flanked by loxP sites.