This knock-out mutant of the Ngly1 gene was generated by crossing mice carrying the Ngly1tm2b(KOMP)Wtsi knock-out/reporter allele to B6N.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/J (JAX Stock#016226) mice and has exon 2 deleted. These mice may be useful for studying proteasome-mediated degradation of misfolded glycoproteins and protein processing in the endoplasmic reticulum.
Cathleen Lutz, The Jackson Laboratory
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Ngly1 | N-glycanase 1 |
The N-glycanase 1 (Ngly1) enzyme deglycosylates the denatured form of N-linked glycoproteins in the cytoplasm and assists their proteasome-mediated degradation. Ngly1 may play a role in the proteasome-mediated degradation of misfolded glycoproteins and protein processing in the endoplasmic reticulum. Diseases associated with NGLY1 include congenital disorder of glycosylation [type iv] and alacrimia-choreoathetosis-liver dysfunction syndrome.
These Ngly1 knock-out mice carry an allele in which exon 2 has been excised. As the mice are characterized, we will modify the strain description and add phenotype data. Homozygotes are viable and fertile. However, pups produced by homozygous females do not survive past postnatal day 1 (no milk spot is observed).
This knock-out mutant of the Ngly1 gene was generated by crossing mice carrying the Ngly1tm2a(KOMP)Wtsi knockout-first allele (JAX Stock#27582) to B6N.Cg-Tg(Sox2-cre)1Amc/J (JAX Stock#014094) mice and then to B6N.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/J (JAX Stock#016226) mice.
The "knockout first, promoterless" Ngly1tm2a(KOMP)Wtsi allele, the L1L2_gt2 cassette was placed upstream of the critical exon (exon 2) of the N-glycanase 1 locus (Ngly1); insertion at bp position Chr14:16,254,154 [Build GRCm38]. The L1L2_gt2 cassette is composed of an FRT site, a mouse En2 intron/splice-acceptor/exon (En2 SA), a self-cleaving 2A peptide, the β-galactosidase gene (lacZ), a second self-cleaving 2A peptide, the neomycin-resistance gene (neo), an SV40 polyadenylation signal (SV40pA), a second FRT site and a loxP site. A second loxP site was inserted downstream of exon 2 at bp position Chr14:16,255,007 bp [Build GRCm38]; therefore flanking exon 2 with loxP sites.
The targeting vector PGS00036_A_C09 was introduced into C57BL/6N-derived JM8.F6 embryonic stem (ES) cells. Correctly targeted ES cell clone EPD0044_5_F07 from the International Knockout Mouse Consortium/International Mouse Phenotyping Consortium was injected into recipient blastocysts. The resulting chimeric animals were tested for germline transmission. Sperm was cryopreserved. Upon arrival at The Jackson Laboratory, the frozen sperm was used to fertilize oocytes from C57BL/6NJ inbred females (Stock No. 005304). Mice carrying the Ngly1tm2a(KOMP)Wtsi allele were bred to B6N.Cg-Tg(Sox2-cre)1Amc/J (Stock No. 014094), generating the knock-out/lacZ reporter Ngly1tm2b(KOMP)Wtsi allele. Mice carrying the Ngly1tm2b(KOMP)Wtsi allele were crossed to B6N.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/J (JAX Stock#016226) mice to excise the FRT flanked lacZ/neo cassette.
Allele Name | targeted mutation 2d, Wellcome Trust Sanger Institute |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | |
Gene Symbol and Name | Ngly1, N-glycanase 1 |
Gene Synonym(s) | |
Strain of Origin | C57BL/6N |
Chromosome | 14 |
Molecular Note | The L1L2_gt2 cassette was placed upstream of the critical exon (exon 2) of the gene; inserting at bp position Chr14:16,254,154 [Build GRCm38]. The cassette is composed of composed of an FRT site, a mouse En2 intron/splice-acceptor/exon (En2 SA), a self-cleaving 2A peptide, the beta-galactosidase gene (lacZ), a second self-cleaving 2A peptide, the neomycin-resistance gene (neo), an SV40 polyadenylation signal (SV40pA), a second FRT site and a loxP site. A second loxP site was inserted downstream of exon 2 at bp position Chr14:16,255,007 bp [Build GRCm38]; thereby flanking exon 2 with loxP sites. Cre-mediated recombination removed exon 2. Flp-mediated recombination removed the FRT-flanked neo cassette |
When maintaining a live colony, heterozygous female mice may be bred to homozygous male mice. Homozygotes are viable and fertile. However, pups produced by homozygous females do not survive past postnatal day 1 (no milk spot is observed).
When using the Ngly1 KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #028975 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Ngly1<tm2d(KOMP)Wtsi> |
Frozen Mouse Embryo | B6N(Cg)-Ngly1<tm2d(KOMP)Wtsi>/LutzyJ Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6N(Cg)-Ngly1<tm2d(KOMP)Wtsi>/LutzyJ Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6N(Cg)-Ngly1<tm2d(KOMP)Wtsi>/LutzyJ Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6N(Cg)-Ngly1<tm2d(KOMP)Wtsi>/LutzyJ Frozen Embryo | $3373.50 |
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