Overview

Also Known As:TDP-43^M337V BAC knock-in

TDP-43^M337V BAC allele is a Gt(ROSA)26Sor knock-in of a loxP-flanked, full-length human TARDBP genomic locus with M337V mutation in exon 6 and C-terminal fluorescent tag. Prior to Cre recombinase exposure, TDP-43^M337V BAC mice have stable, single-copy, low-level expression of TDP-43^M337V-Ypet that recapitulates many of the key features of ALS/FTD (amyotrophic lateral sclerosis [Lou Gehrig's disease] / frontotemporal dementia [frontotemporal lobar degeneration]), including progressive motor deficits in conjunction with neuromuscular junction abnormalities and reduced survival.

Donating Investigator

Kevin Talbot, University of Oxford

Genetic overview

Genetic Background	Generation

Gt(ROSA)26Sor^{tm1(TARDBPM337V/Ypet)Tlbt}*

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence, Humanized sequence)	Gt(ROSA)26Sor	gene trap ROSA 26, Philippe Soriano

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Research Applications

Research Tools
Neurobiology Research

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

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) and frontotemporal dementia (FTD or frontotemporal lobar degeneration). The TDP-43 methionine-to-valine (M337V) missense mutation is associated with increased low molecular weight aggregates and neuronal dysfunction.

The TDP-43^M337V BAC allele has a loxP-flanked, full-length human TARDBP genomic locus with M337V mutation in exon 6 and C-terminal fluorescent tag, all inserted into the Gt(ROSA)26Sor locus. Exposure to Cre recombinase leads to deletion of the TDP-43^M337V BAC sequences. The phenotype of TDP-43^M337V BAC mice prior to Cre recombinase is described below.

In contrast to transgene overexpression models, this Gt(ROSA)26Sor knock-in TDP-43^M337V BAC allele exhibits stable, single-copy, low-level expression of TDP-43^M337V-Ypet driven by the endogenous human promoter. Both homozygous and heterozygous mice of both sexes recapitulate many of the key features of ALS/FTD; including progressive motor deficits in conjunction with neuromuscular junction (NMJ) abnormalities and reduced survival.

At 12 months of age, despite a clear neuromuscular phenotype, homozygotes and heterozygotes do not demonstrate cytoplasmic mislocalization/accumulation of TDP-43 - the key pathological hallmark of ALS in the CNS.

Reduced survival is observed in both sexes (homozygotes ~500 days; heterozygotes ~650 days). The phenotype is dose-dependent: homozygous mice have an earlier age-of-onset and are more severely affected than heterozygous littermates.
In addition, cultured spinal cord motor neurons derived from heterozygous embryos show a characteristic phenotype of reduced survival, translocation of TDP-43 from nucleus to cytoplasm and reduced stress granule formation.

Homozygous and heterozygous mice are viable and fertile, with no apparent deficits in fertility or litter size compared to controls. Human TDP-43^M337V-Ypet is expressed at low levels in all tissues examined in the expected pattern of cellular distribution, with no significant expression level differences reported between heterozygous or homozygous mice (cortex and spinal cord). In heterozygous mice, the TDP-43^M337V-Ypet expression level is ~40% of endogenous levels.

The donating investigator reports that C-terminal Ypet tag is suitable for immunostaining via antibody directed against GFP, but not for direct fluorescence.

Development

The TDP-43^M337V BAC mice were created by collaboration led by Dr. Kevin Talbot (University of Oxford). BAC cloning was performed in the laboratory of Dr. Richard Wade-Martins (University of Oxford), using the RP11-829B14 BAC as source DNA for the full-length human TARDBP genomic locus. The human TARDBP was then modified by site-directed mutagenesis to create the M337V missense mutation in exon 6 (an A-to-G base change [A1009G] encoding a methionine-to-valine substitution at amino acid position 337). The final TDP-43-M337V-Ypet BAC construct includes the TDP-43-M337V genomic DNA (44,886 bp) with a Ypet ORF cDNA sequence (717 bp; described below) fused to the 5' end of TARDBP exon 6, as well as 9.2 kbp of the original BAC backbone sequences. The TDP-43-M337V-Ypet BAC sequences are flanked with loxP sites. In the mouse core facility directed by Dr. Ben Davies (Wellcome Trust Centre for Human Genetics, University of Oxford), (C57BL/6J x 129S6/SvEvTac)F1-derived IDG26.10-3 embryonic stem (ES) cells with a pre-integrated Gt(ROSA)26Sor locus (splice acceptor::frt::PGK promoter-attP-hygromycin resistance gene-pA::frt::attP between exons 1 and 2) were re-transfected with the TDP-43-M337V-Ypet BAC sequence and a PhiC31 vector for integrase-mediated cassette exchange. The correctly re-targeted ES cells, with the Gt(ROSA)26Sor containing (from 5' to 3') splice acceptor::frt::PGK promoter-attR-neomycin resistance gene-bGHpA::loxP::TDP-43-M337V-Ypet BAC sequence::loxP::attL, were injected into recipient blastocysts. Chimeric males were bred with C57BL/6J females to establish germline transmission. The resulting TDP-43^M337V BAC colony was additionally backcrossed to C57BL/6J wildtype mice for three generations prior to sending homozygous males with black coat color to The Jackson Laboratory Repository in 2016. 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).

Of note, the donating investigator reports that the Y chromosome may not have been fixed to the C57BL/6J background during backcrossing.

The Ypet cDNA sequence used here is described in Nguyen and Daugherty 2005 Nat Biotechnol. 23:355. The codon-improved YFP variant YFP3 (with F46L, I47L, S208F, V224L, H231E, D234N mutations) was further modified to include several Venus mutations (F64L, R79K, M153T, V163A, S175G), resulting in the YFP for energy transfer (YPet) variant.

Expression Data

Expressed Gene	TARDBP, TAR DNA binding protein, human
Expressed Gene	YFP, Yellow Fluorescent Protein, jellyfish
Site of Expression	When these mutant mice are bred to mice that express Cre recombinase, the resulting offspring will express YFP (revealed by immunostaining via antibody directed against GFP) in the cre-expressing tissues.

Control Suggestions

Wild-type from the colony

Approximate Controls

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Gordon D; Dafinca R; Scaber J; Alegre-Abarrategui J; Farrimond L; Scott C; Biggs D; Kent L; Oliver PL; Davies B; Ansorge O; Wade-Martins R; Talbot K. 2018. Single-copy expression of an amyotrophic lateral sclerosis-linked TDP-43 mutation (M337V) in BAC transgenic mice leads to altered stress granule dynamics and progressive motor dysfunction. Neurobiol Dis 121:148-162PubMed: 30290270 MGI: J:265544

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Genetics

Gt(ROSA)26Sor^{tm1(TARDBPM337V/Ypet)Tlbt}*

Allele Symbol: Gt(ROSA)26Sor^{tm1(TARDBPM337V/Ypet)Tlbt}*

Allele Name	targeted mutation 1, Kevin Talbot
Allele Type	Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence, Humanized sequence)
Allele Synonym(s)	TDP-43^M337V
Gene Symbol and Name	Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano
Gene Synonym(s)
Expressed Gene	TARDBP, TAR DNA binding protein, human
Expressed Gene	YFP, Yellow Fluorescent Protein, jellyfish
Site of Expression	When these mutant mice are bred to mice that express Cre recombinase, the resulting offspring will express YFP (revealed by immunostaining via antibody directed against GFP) in the cre-expressing tissues.
Strain of Origin	(C57BL/6J x 129S6/SvEvTac)F1
Chromosome	6
General Note	embryonic stem cell line = IDG26.10-3
Molecular Note	The human TARDBP from BAC RP11-829B14 was modified by site-directed mutagenesis to create the M337V missense mutation in exon 6 (an A-to-G base change (A1009G) encoding a methionine-to-valine substitution at amino acid position 337). The final construct includes the TDP-43-M337V genomic DNA (44,886 bp) with a Ypet ORF cDNA sequence (717 bp; described below) fused to the 5' end of TARDBP exon 6, as well as 9.2 kbp of the original BAC backbone sequences. The TDP-43-M337V-Ypet BAC sequences are flanked with loxP sites. Embryonic stem (ES) cells with a pre-integrated Gt(ROSA)26Sor locus (splice acceptor::FRT::PGK promoter-attP-hygromycin resistance gene-pA::FRT::attP between exons 1 and 2) were re-transfected with the TDP-43-M337V-Ypet BAC sequence and a PhiC31 vector for integrase-mediated cassette exchange. The Ypet cDNA sequence is a codon-improved YFP variant YFP3 (with F46L, I47L, S208F, V224L, H231E, D234N mutations) further modified to include several Venus mutations (F64L, R79K, M153T, V163A, S175G), resulting in the YFP for energy transfer (YPet) variant.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Research Tools
- Genetics Research
  - Mutagenesis and Transgenesis: Cre-lox System
  - Tissue/Cell Markers: Cre-lox System
- Fluorescent Proteins
- Cre-lox System
  - loxP-flanked Sequences
- Developmental Biology Research
  - Cre-lox System
- Neurobiology Research
  - cell marker
Neurobiology Research
- Myelination Defects
  - peripheral neuropathy
- Amyotrophic Lateral Sclerosis (ALS)
- Ataxia (Movement) Defects
- Neurodegeneration
- Neuromuscular defects
- Cre-lox System
  - loxP-flanked Sequences
- Fluorescent protein expression in neural tissue

Mammalian Phenotype Terms by Genotype

References

Gordon D; Dafinca R; Scaber J; Alegre-Abarrategui J; Farrimond L; Scott C; Biggs D; Kent L; Oliver PL; Davies B; Ansorge O; Wade-Martins R; Talbot K. 2018. Single-copy expression of an amyotrophic lateral sclerosis-linked TDP-43 mutation (M337V) in BAC transgenic mice leads to altered stress granule dynamics and progressive motor dysfunction. Neurobiol Dis 121:148-162PubMed: 30290270 MGI: J:265544

Additional - Gt(ROSA)26Sor^{tm1(TARDBPM337V/Ypet)Tlbt}* related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Sanger sequencing:Gt(ROSA)26Sor SEQ Alt1
- Standard PCR:Gt(ROSA)26Sor-Alternate 1
- Genotyping resources and troubleshooting
Breeding Considerations

Homozygous and heterozygous mice are viable past one year of age and fertile, with no apparent deficits in fertility or litter size compared to controls. When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). Alternatively, homozygous mice may be bred together.
- Additional Breeding and Husbandry Support
Citation
When using the TDP-43^M337V BAC knock-in mouse strain in a publication, please cite the originating article(s) and include JAX stock #029266 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

Cryo Recovery

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Heterozygous for Gt(ROSA)26Sor<tm1(TARDBP*M337V/Ypet)Tlbt>

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Also Known As:TDP-43M337V BAC knock-in

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Gt(ROSA)26Sortm1(TARDBP*M337V/Ypet)Tlbt

Allele Symbol: Gt(ROSA)26Sortm1(TARDBP*M337V/Ypet)Tlbt

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

References

Additional - Gt(ROSA)26Sortm1(TARDBP*M337V/Ypet)Tlbt related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Also Known As:TDP-43^M337V BAC knock-in

Gt(ROSA)26Sor^{tm1(TARDBPM337V/Ypet)Tlbt}*

Allele Symbol: Gt(ROSA)26Sor^{tm1(TARDBPM337V/Ypet)Tlbt}*

Additional - Gt(ROSA)26Sor^{tm1(TARDBPM337V/Ypet)Tlbt}* related