Overview

Also Known As:amiR-TDP43i

These amiR-TDP43i (or CAG-RFP-amiR-TDP43i) transgenic mice generally express a red fluorescent protein (dsRed2) and an artificial miRNA (amiR-TDP43) that targets the mouse TAR DNA binding protein mRNA for knockdown. Transgenic mice have reduced TDP-43 expression resulting in a rapid, progressive neurodegenerative phenotype that recapitulates many features of human amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD).

Donating Investigator

Zuoshang Xu, University of Massachusetts Medical School

Genetic overview

Genetic Background	Generation

Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu

Allele Type
Transgenic (Reporter)

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Research Applications

Research Tools
Neurobiology Research
Cell Biology Research
Mouse/Human Gene Homologs

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

These amiR-TDP43i (or CAG-RFP-amiR-TDP43i) transgenic mice have widespread expression of both red fluorescent protein (dsRed2) and an artificial miRNA (amiR-TDP43) that targets the mouse TAR DNA binding protein mRNA for knockdown. Expression is directed by the CMV-IE enhancer/chicken β-actin/rabbit β-globin hybrid promoter. amiR-TDP43 expression in the central nervous system and periphery (including brain [both neurons and glia], spinal cord, muscle, liver and other organs) results in a modest reduction (~10-20%) of TDP-43 expression in central nervous system homogenates. dsRed2 expression may be observed by direct/native fluorescence, by mRNA (in situ) hybridization, and by antibody staining/immunohistochemistry. DsRed2 excitation and emission maxima are 563nm and 582nm, respectively. In the cortex, dsRed2 immunofluorescence is observed in ~30-40% of neurons, astrocytes and oligodendrocytes, but not in microglia. As expected, no GFP expression is reported in brain or spinal cord.

amiR–TDP43i hemizygotes are viable with no gross physical abnormalities at birth. Hemizygous mice may begin to display progressive neurological defects around the time they are weaned, including head shaking/bobbing, hyperactivity (~4-5 weeks), weight loss (~5 weeks), circling, wobbly gait (~6-7 weeks), hypoactivity (~10 weeks), paralysis (~9-20 weeks) and death by 9-20 weeks. amiR–TDP43i hemizygotes display three end-stage phenotypes: hindlimb paralysis, sudden death, and whole-body paralysis. Muscle denervation is evident by electromyography shortly after weaning. Progressive motor neuron degeneration results in the end disease stage, at which there is ~25% loss of layer 5 cortical neurons and ~60% loss of lumbar spinal cord ventral horn ChAT+ motor neurons. Many transgenic mice exhibit heart enlargement. The average lifespan is slightly shorter for males than females, but the difference is not statistically significant. The donating investigator has not attempted to generate homozygous mice to date (December 2012), and reports transgenic males breed better than transgenic females. Additionally, because the phenotype may become more severe if the mice are backcrossed with FVB/NJ for two or more generations, the colony is maintained at The Jackson Laboratory Repository by breeding transgenic males with (C57BL/6J x FVB/NJ)F1 hybrid females (Stock No. 019019) every generation.

Of note, the neuromuscular-phenotype-free, cre-inducible parental strain used to generate the amiR-TDP43i transgenic mice is also available at The Jackson Laboratory as Stock No. 017919 (amiR-TDP43u line 6 or CAG-loxp-EGFP/3xpA-loxp-RFP-amiR-TDP43).

Development

The CAG::loxP::EGFP/3xpA::loxP::dsRed2::intron::amiR-TDP43::intron::polyA transgene (or amiR-TDP43u transgene) was designed in the laboratory of Dr. Zuoshang Xu (University of Massachusetts Medical School). This transgene contained (from 5' to 3') the CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from the pCAGG vector), a loxP site, an enhanced green fluorescent protein sequence (from pEGFP-N1) followed by three polyadenylation signals, a second loxP site, a dsRed2 fluorescent protein sequence (from pDsRed2-C1), a synthetic artificial intron containing the amiR-TDP43 precursor miRNA sequence, and a polyA signal. The synthetic artificial intron utilized here was designed to contain typical intron splicing regulatory elements and was specifically placed 10 nucleotides downstream of the RFP stop codon because introns within 50 nucleotides downstream from the stop codon are found in some eukaryotic mRNAs and do not cause non-sense mediated decay (NMD). The amiR-TDP43 sequence (miR-TDP-43b; 5'-ATGCTGAACCTAAGCATAA-3') is an artificial precursor miRNA sequence encoding a short hairpin RNA specifically synthesized to target the RNA of mouse TAR DNA binding protein (TBP-43 or Tardbp). The resulting transgene was injected into (C57BL/6 x SJL)F2 hybrid mouse embryos. Transgenic mice from founder line 6 were identified, and bred with (C57BL/6J x FVB/NJ)F1 mice to establish germline transmission of the transgene. The amiR-TDP43u line 6 transgenic males (see Stock No. 017919) were then bred to mice with widespread expression of Cre recombinase (CMV-Cre on FVB genetic background) to remove the floxed EGFP sequences. The resulting amiR-TDP43i transgenic mice (line 6-50) were subsequently bred to (C57BL/6J x FVB/NJ)F1 female mice every generation for eight generations prior to sending to The Jackson Laboratory Repository. The donating investigator reports the CMV-Cre transgene was removed after two generations, and they selected breeders by high RFP fluorescence in tail samples. The donating investigator also reports amiR-TDP43i transgenic mice have two copies of the transgene inserted in head-to-head orientation on chromosome 14 band A2. Upon arrival at The Jackson Laboratory Repository, transgenic males were bred to (C57BL/6J x FVB/NJ)F1 hybrid females (Stock No. Stock No. 019019) every generation to maintain the colony.

Expression Data

Expressed Gene	RFP, Red Fluorescent Protein, coral
Site of Expression	Widespread expression of both red fluorescent protein (dsRed2) and an artificial miRNA (amiR-TDP43). Expression of amiR-TDP43 in the central nervous system and periphery (including brain [both neurons and glia], spinal cord, muscle, liver and other organs) results in a modest reduction (~10-20%) of TDP-43 (Tardbp) expression in central nervous system homogenates.

Control Suggestions

The genetic background control is (C57BL/6J x FVB/NJ)F1 hybrid mice (Stock No. 019019). This F1 is created by breeding C57BL/6J females (Stock No. 000664) with FVB/NJ males (Stock No. 001800).
Noncarrier

Additional Information

Considerations for Choosing Controls

Selected References

Yang C; Wang H; Qiao T; Yang B; Aliaga L; Qiu L; Tan W; Salameh J; McKenna-Yasek DM; Smith T; Peng L; Moore MJ; Brown RH Jr; Cai H; Xu Z. 2014. Partial loss of TDP-43 function causes phenotypes of amyotrophic lateral sclerosis. Proc Natl Acad Sci U S A 111(12):E1121-9PubMed: 24616503 MGI: J:207750

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Genetics

Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu

Allele Symbol: Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu

Allele Name	transgene insertion 6, Zuoshang Xu
Allele Type	Transgenic (Reporter)
Allele Synonym(s)	amiR-TDP43i
Gene Symbol and Name	Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu, transgene insertion 6, Zuoshang Xu
Gene Synonym(s)
Promoter	CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter,
Expressed Gene	RFP, Red Fluorescent Protein, coral
Site of Expression	Widespread expression of both red fluorescent protein (dsRed2) and an artificial miRNA (amiR-TDP43). Expression of amiR-TDP43 in the central nervous system and periphery (including brain [both neurons and glia], spinal cord, muscle, liver and other organs) results in a modest reduction (~10-20%) of TDP-43 (Tardbp) expression in central nervous system homogenates.
Strain of Origin	(C57BL/6 x SJL)F2
Chromosome	UN
Molecular Note	The transgenic construct contains (from 5' to 3') the CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, a loxP site, an enhanced green fluorescent protein sequence followed by three polyadenylation signals, a second loxP site, a dsRed2 fluorescent protein sequence, a synthetic artificial intron containing the amiR-TDP43 precursor miRNA sequence, and a polyA signal. The synthetic artificial intron utilized here was designed to contain typical intron splicing regulatory elements and was specifically placed 10 nucleotides downstream of the RFP stop codon because introns within 50 nucleotides downstream from the stop codon are found in some eukaryotic mRNAs and do not cause non-sense mediated decay (NMD). The amiR-TDP43 sequence (miR-TDP-43b; 5'-ATGCTGAACCTAAGCATAA-3') is an artificial precursor miRNA sequence encoding a short hairpin RNA specifically synthesized to target the RNA of mouse TAR DNA binding protein (TBP-43 or Tardbp).

Disease/Phenotype

Disease Terms

Characteristics of this human disease are associated with transgenes and other mutation types in the mouse.

amyotrophic lateral sclerosis type 10

Research Areas By Phenotype

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

Research Tools
- Neurobiology Research
  - cell marker
- Genetics Research
  - Mutagenesis and Transgenesis: transcriptional activation
  - Tissue/Cell Markers: astrocytes, neurons
  - Tissue/Cell Markers: neurons
- Fluorescent Proteins
Neurobiology Research
- Fluorescent protein expression in neural tissue
- Cortical Defects
- Neuromuscular defects
- Myelination Defects
  - peripheral neuropathy
- Amyotrophic Lateral Sclerosis (ALS)
- Astrocyte Defects
- Ataxia (Movement) Defects
- Neurodegeneration
Cell Biology Research
- Transcriptional Regulation
Mouse/Human Gene Homologs
- amyotrophic lateral sclerosis (ALS)

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu/0
involves: C57BL/6 * C57BL/6J * FVB/NJ * SJL

behavior/neurological phenotype

abnormal gait
- unsteady gait

decreased vertical activity
- mice show reduced vertical behaviors by 5 weeks of age

head bobbing

hindlimb paralysis
- 17.4% of males, and no females, have a short lifespan and die with hindlimb paralysis

hyperactivity
- mice exhibit hyperactivity beginning at 4 weeks of age, which peaks at around 5 weeks of age and ends around 10 weeks of age

hypoactivity
- after 10 weeks of age, mice are less active as they become progressively weaker

impaired limb coordination
- discoordinated movements

paralysis
- mice surviving the longest develop whole-body paralysis; this is seen in both males and females

cellular phenotype

skeletal muscle fiber necrosis
- necrotic skeletal muscle fibers and fibers with centralized nuclei in some areas of the muscle

growth/size/body region phenotype

enlarged heart

weight loss
- mice start to lose weight around 5 weeks of age

hematopoietic system phenotype

microgliosis
- abundant microgliosis in the ventral horn gray matter

immune system phenotype

microgliosis
- abundant microgliosis in the ventral horn gray matter

mortality/aging

premature death
- 17.4% of males, and no females, have a short lifespan and die with hindlimb paralysis at a mean of 38.9 days
- 25% of males and 15.6% of females show sudden death without paralysis with a mean of 33.6 days and 41.3 days in females and males, respectively
- 57.6% of males and 84.4% of females survive longer (mean of 80-83 days) and die with whole body paralysis

muscle phenotype

abnormal muscle morphology
- necrotic skeletal muscle fibers and fibers with centralized nuclei in some areas of the muscle

muscle degeneration
- electromyography indicates that mice develop progressive active denervation of muscles as indicated by the presence of positive sharp waves

progressive muscle weakness

skeletal muscle fiber necrosis
- necrotic skeletal muscle fibers and fibers with centralized nuclei in some areas of the muscle

nervous system phenotype

astrocytosis
- abundant astrogliosis in the ventral horn gray matter
- astrogliosis in the deep layer of the frontal cortex

axon degeneration
- degeneration of axons in the ventral but not dorsal roots

microgliosis
- abundant microgliosis in the ventral horn gray matter

motor neuron degeneration
- dramatic motor neuron loss, with about 60% fewer motor neurons in the ventral horn

neurodegeneration
- neurodegeneration in layer V cortex and ventral horn spinal cord

cardiovascular system phenotype

enlarged heart

References

Yang C; Wang H; Qiao T; Yang B; Aliaga L; Qiu L; Tan W; Salameh J; McKenna-Yasek DM; Smith T; Peng L; Moore MJ; Brown RH Jr; Cai H; Xu Z. 2014. Partial loss of TDP-43 function causes phenotypes of amyotrophic lateral sclerosis. Proc Natl Acad Sci U S A 111(12):E1121-9PubMed: 24616503 MGI: J:207750

Additional - Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Tg(CAG-dsRed2/miRTDP43)6-50Zxu
- Standard PCR:Tg(CAG-dsRed2/miRTDP43)6-50Zxu
- Genotyping resources and troubleshooting
Breeding Considerations

The donating investigator has not attempted to generate homozygous mice to date (December 2012). The donating investigator reports transgenic males breed better than transgenic females. In addition, the neurodegenerative phenotype may become more severe if backcrossed with FVB/NJ for two or more generations. As such, the colony is maintained at The Jackson Laboratory Repository by breeding transgenic males with (C57BL/6J x FVB/NJ)F1 hybrid females (Stock No. 019019) every generation.
- Additional Breeding and Husbandry Support
Citation
When using the amiR-TDP43i mouse strain in a publication, please cite the originating article(s) and include JAX stock #017934 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 - Not-For-Profit & Academic Pricing

Service/Product	Description	Price
	Hemizygous or Non carrier for Tg(CAG-dsRed2/miRTDP43)6Zxu

Related Products and Services

Frozen Mouse Embryo	STOCK Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu/J	$2595.00
Frozen Mouse Embryo	STOCK Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu/J	$2595.00
Frozen Mouse Embryo	STOCK Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu/J	$3373.50
Frozen Mouse Embryo	STOCK Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu/J	$3373.50

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Also Known As:amiR-TDP43i

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu

Allele Symbol: Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu

Disease Terms

Characteristics of this human disease are associated with transgenes and other mutation types in the mouse.

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu/0involves: C57BL/6 * C57BL/6J * FVB/NJ * SJL

behavior/neurological phenotype

cellular phenotype

growth/size/body region phenotype

hematopoietic system phenotype

immune system phenotype

mortality/aging

muscle phenotype

nervous system phenotype

cardiovascular system phenotype

References

Additional - Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu 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 - Not-For-Profit & Academic Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Genotype: Tg(CAG-dsRed2/RNAi:Tardbp)6Zxu/0
involves: C57BL/6 * C57BL/6J * FVB/NJ * SJL