B6J.B6N-Tardbp^{tm1.1(TARDBP)Mdk}/J

Stock number: 039101
Product name: TARDBP-GR wt
Strain synonyms: TARDBP-GR
Research areas: Mouse/Human Gene Homologs, Neurobiology Research, Research Tools

Description

Humanized TARDBP-GR (gene replacement) mice are a knock-in/knock-out model in which the mouse genomic region from Tardbp (TAR DNA binding protein) is exchanged for the 65 kb human TARDBP genomic region. These mice may serve as base models into which additional alterations can be introduced to explore more fully the role of TARDBP in frontotemporal lobar degeneration (FTLD-TDP) and dementias.

Detailed description

The human TAR DNA binding protein (TARDBP or TDP-43) is an ubiquinated, DNA/RNA-binding protein localized to the nucleus of cells. TDP-43 mutations in the amyloidogenic core region (residues 311-360) and the glycine-rich, flexible C-terminal region are associated with the development of amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease), frontotemporal lobar degeneration (FTLD-TDP) and as a risk variant for Alzheimer's disease.

These humanized TARDBP-GR (gene replacement) knock-in/knock-out mice carry a 65 kb fragment of human TARDPB replacing the mouse Tardbp locus. Mice homoozygous for the mutation are viable and fertile with no obvious phenotype. These mice may be useful for studying TARDBP function.

Development

The targeting vector was designed to insert a 65 kb (Chr1:10,989,074-11,054,422) fragment of the human TARDBP (TAR DNA binding protein) in place of Chr4:148,679,229-148,723,697 of the mouse Tardbp gene on chromosome 4. The genomic sequences are from human assembly GRCh38.p14 and the mouse assembly GRCm39. The construct included a loxP-flanked neomycin selection cassette. In addition, there is also a TARDBP non-coding antisense RNA in the human genome segment. The insertion disrupts Tardbp transcription. The construct was electroporated into C57BL/6N-PRX-B6N embryonic stem cells (ES). The neomycin cassette was removed by transient expression of Cre. Correctly targeted ES cells were injected into blastocysts. Chimeric animals were backcrossed to C57BL/6J mice for at least 5 generations prior to sending males to The Jackson Laboratory Repository in 2024. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

In 2024, 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 was segregating, the Thy1 marker on Chromosome 9.

Allele

Symbol: Tardbp^{tm1.1(TARDBP)Mdk}
Name: targeted mutation 1.1, Michael D Koob
MGI accession ID: MGI:7576944
Generation method: Targeted
Attributes: Null/Knockout; Inserted expressed sequence; Humanized sequence
Synonyms: TARDBP-GR
Marker symbol: Tardbp
Marker name: TAR DNA binding protein
Marker synonyms: 1190002A23Rik; ALS10; Tdp43; TDP-43
Chromosome: 4
Strain of origin: C57BL/6NTac
Molecular note: The targeting vector is designed to insert a 65 kb (Chr1: 10989074 - 11054422) fragment of the human TARDBP (TAR DNA binding protein) in place of Chr4:148679229 - 148723697 of the mouse Tardbp gene. The genomic sequences are from human assembly GRCh38.p14 and the mouse assembly GRCm39. The insertion disrupts Tardbp transcription. The construct includes an loxP-flanked neomycin selection cassette. In addition, there is also a TARDBP (non-coding antisense RNA) in the human genome segment. Transient cre-mediated recombination removed the neo cassette.