STOCK Nde1^tm1Caw/YfngJ

Stock number: 038493
Product name: Nde1^-
Research areas: Cell Biology Research, Developmental Biology Research, Neurobiology Research, Research Tools

Description

Nde1^-/- mice have exon 2 of the nudE neurodevelopment protein 1 gene removed, abolishing expression. These mice may be useful for studying microtubule organization, mitosis, neuronal migration, and cerebral cortex size.

Detailed description

Nde1^-/- mice have exon 2 of the nudE neurodevelopment protein 1 (Nde1) gene removed, abolishing expression. NDE1 is highly expressed in the developing cerebral cortex in cortical progenitors and young post-mitotic neurons where it is required for proper cell cycle progression, mitosis, chromatin remodeling, and genome stability.

Homozygous Nde1^-/- mice are viable and fertile and have a smaller brain and smaller cerebral cortex with dramatically reduced superficial cortical layers when compared to wildtype mice. They show little anatomical changes in other brain structures such as the hippocampus, striatum, and cerebellum. The loss of superficial cortical neurons in homozygous mutants results from Trp53-dependent apoptosis in response to DNA double stand break.

Development

The Nde1^- targeting vector was designed to insert a loxP site upstream of exon 2 and a loxP-flanked neo cassette downstream of exon 2 of the nudE neurodevelopment protein 1 (Nde1) gene. The construct was electroporated into 129S4/SvEvTac-derived J1 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a Cre recombinase expression plasmid to remove the neo cassette and exon 2. Resulting clones lacking the neo cassette and exon 2 were injected into C57BL/6 blastocysts. The resulting chimeric males were crossed to Black Swiss females for germline transmission. Resulting Nde1^-/- mice were backcrossed to C57BL/6 mice for five generations by the donating laboratory (see SNP note below). 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).

In 2023, 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. Eight of the 43 markers throughout the genome were segregating with 129 and an unknown source suggesting an incomplete backcross.

Allele

Symbol: Nde1^tm1Caw
Name: targeted mutation 1, Christopher A Walsh
MGI accession ID: MGI:3514026
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: Nde1^-
Marker symbol: Nde1
Marker name: nudE neurodevelopment protein 1
Marker synonyms: 2810027M15Rik; HOM-TES-87; LIS4; MHAC; mNudE; NDE; Nude; NUDE1
Chromosome: 16
Strain of origin: 129S4/SvJae
Molecular note: A loxP site was inserted upstream of exon 2 and a floxed PGK-neomycin cassette was inserted downstream. Transient transfection of properly targeted ES cells with a Cre-expressing plasmid resulted in excision of the selection cassette together with exon 2. Immunoblot analysis confirmed the absence of protein product.