C57BL/6J-Ngly1^em10Lutzy/J

Stock number: 028221
Product name: Ngly1^R539X
Strain synonyms: C57BL/6J-Ngly1<em10(R539X)Lutzy>/J
Research areas: Developmental Biology Research, Neurobiology Research, Research Tools

Description

The Ngly1^R539X allele has a 1 bp replacement in the mouse N-glycanase 1 gene that encodes a nonsense mutation, resulting in a premature stop codon. This mutation corresponds to the R542X mutation identified in the patient population as a disease causing variant in Congenital Disorders of Glycosylation Type 1v.

Detailed description

The Ngly1^R539X allele has a 1 bp replacement (CGA-to-TGA) in the mouse N-glycanase 1 gene that encodes an R539X nonsense mutation, resulting in a premature stop codon. This mutation corresponds to the R542X mutation in human patients identified as a disease causing variant in Congenital Disorders of Glycosylation Type 1v; an autosomal recessive multi-system disorder characterized by global developmental delay, hypotonia, abnormal involuntary movements and poor tear production. N-glycanase 1 is an enzyme that catalyzes the deglycosylation of misfolded N-linked glycoproteins so that they can be processed properly. Mice heterozygous for the Ngly1^R539X allele are viable and fertile. Homozygous mice are lethal. The donating investigator reports homozygotes are present in Mendelian ratios to embryonic day (E)18, and ~20% of the expected homozygotes are born but die within 1-2 days.
To date (September 2015), full characterization is in progress. As the mice are characterized, we will modify the strain description.

This mouse model is available by way of a collaborative effort between the Grace Wilsey Foundation and The Jackson Laboratory.

Development

The Ngly1^R539X allele was created by Dr. Cathleen Lutz (The Jackson Laboratory) using CRISPR/cas9 endonuclease-mediated genome editing in C57BL/6J mouse zygotes (Stock No. 000664).
Specifically, the N-glycanase 1 locus (Ngly1) on chromosome 14 was targeted with single guide RNAs designed to create a C-to-T nucleotide replacement (CGA-to-TGA) encoding an arginine 539-to-termination codon nonsense mutation (R539X). The sgRNAs and Cas9 endonuclease were introduced into C57BL/6J-derived fertilized eggs by pronuclear injection. Embryos were transferred to pseudopregnant females, and correctly targeted pups (identified by sequencing and PCR) were bred to C57BL/6J inbred mice for germline transmission. Mice from founder 93 were further bred to C57BL/6J inbred mice to develop the colony.

Allele

Symbol: Ngly1^em10Lutzy
Name: endonuclease-mediated mutation 10, Cathy Lutz
MGI accession ID: MGI:5752820
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Marker symbol: Ngly1
Marker name: N-glycanase 1
Chromosome: 14
Strain of origin: C57BL/6J
Molecular note: Using CRISPR/cas9 endonuclease mediated genome editing, single guide RNAs are designed to create a C-to-T nucleotide replacement (CGA-to-TGA) encoding an arginine 539-to-termination codon nonsense mutation (R539X). This mutation corresponds to the R542X mutation identified in the patient population as a disease causing variant in Congenital Disorders of Glycosylation Type 1v.