Overview

Also Known As:MthfrC677T/APOE4/Trem2R47H

Mthfr*C677T/APOE4/Trem2*R47H is a triple mutant strain carrying a CRISPR/Cas9-generated A262V point mutation of the methylenetetrahydrofolate reductase gene that models the human C677T polymorphism linked to risk of Alzheimer's disease, a humanized ApoE knock-in mutation (sequence coding for isoform E4) and a CRISPR/Cas9-generated R47H point mutation of the Trem2 gene. These mice may be suitable for use in studies related to Alzheimer's disease, lipoproteins, arteriosclerosis, and coronary heart disease.

Donating Investigator

Mike Sasner, The Jackson Laboratory

Genetic overview

Genetic Background	Generation

Trem2^em1Adiuj

Allele Type	Gene Symbol	Gene Name
Endonuclease-mediated (Humanized sequence)	Trem2	triggering receptor expressed on myeloid cells 2

Mthfr^em1Adiuj

Allele Type	Gene Symbol	Gene Name
Endonuclease-mediated (Humanized sequence)	Mthfr	methylenetetrahydrofolate reductase

Apoe^{tm1.1(APOE4)Adiuj}*

Allele Type	Gene Symbol	Gene Name
Targeted (Inserted expressed sequence, Humanized sequence)	Apoe	apolipoprotein E

View Genetics

Research Applications

Research Tools
Neurobiology Research
Cardiovascular Research
Metabolism 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

This Mthfr*C677T/APOE4/Trem2*R47H triple mutant strain carries an A262V point mutation of the Mthfr gene that models the human C677T polymorphism, a humanized ApoE knock-in allele, in which exons 2, 3 and most of exon 4 of the mouse Apoe gene were replaced by human APOE4 gene sequence including exons 2, 3 and 4 (and some 3' UTR sequence) and a knock-in of a point mutation into mouse Trem2 gene containing an R47H point mutation with two silent mutations.

The Mthfr A262V point mutation models the human MTHFR C677T polymorphism which is associated with an increased risk for Alzheimer's Disease. The targeted Apoe gene encodes apolipoprotein E, which is important in lipoprotein metabolism and cardiovascular disease as well as Alzheimer's disease, immunoregulation and cognition. The targeted Trem2 gene (triggering receptor expressed on myeloid cells 2) encodes a protein that is part of a receptor signaling complex with TYRO protein tyrosine kinase binding protein, and that activates macrophages and dendritic cells during immune responses. The TREM2 R47H mutation is a missense mutation in exon 2 that is one of the strongest genetic risk factors for late-onset Alzheimer's disease.

Mice that are heterozygous for Mthfr^em1Adiuj (Mthfr*C677T), homozygous for Apoe^{tm1.1(APOE*4)Adiuj} (APOE4) and homozygous for Trem2^em1Adiuj (Trem2*R47H) are viable and fertile [March 2020]. If additional characterization of these Mthfr*C677T/APOE4/Trem2*R47H mice is performed, we may modify the strain description accordingly.

Of note, in brains of mice homozygous for the Trem2^em1Adiuj allele (and not carrying any other mutant alleles), expression of both transcripts of Trem2 is decreased by about 50%. Mice expressing the Trem2 R47H mutation also express a novel splice variant with a deletion of 119bp at the 5' end of exon 2, due to a cryptic splice acceptor site in exon 2 (see Stock No. 027918).

Development

This Mthfr*C677T/APOE4/Trem2*R47H triple mutant line was generated by utilizing CRISPR/Cas9 endonuclease mediated genome editing of the Mthfr gene in B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J (available as Stock No. 028709) mice.

The double mutant B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J (Stock No. 028709) was generated by crossing 2 single mutant lines.

For the Apoe^{tm1.1(APOE*4)Adiuj} allele (APOE4; available as Stock No. 027894): A targeting vector containing a 3' FRT flanked neo cassette was used to replace exons 2, 3 and most of exon 4 with 1.5 kb of human APOE gene sequence including exons 2, 3 and 4 (and some 3' UTR sequence). The human ApoE sequence also contains a 2 nucleotide A to T substitution in intron 3 for identification of the targeted allele by Southern analysis. The construct was electroporated into C57BL/6J derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission by crossing to C57BL/6J. Heterozygous animals were crossed subsequently to FLP recombinase expressing mice (see Stock No. 005703) to remove the FRT site flanked PGK-neo cassette. Mice that no longer contained the FRT flanked PGK-neo cassette were then backcrossed to C57BL/6J at least once to remove the FLP recombinase transgene.

For the Trem2^em1Adiuj allele (Trem2*R47H; available as Stock No. 027918): Plasmids encoding a single guide RNA designed to introduce a R47H point mutation, with two silent mutations, into the Trem2 gene and the cas9 nuclease were introduced into the cytoplasm C57BL/6J-derived fertilized eggs with well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by sequencing and PCR and further bred to C57BL/6J (Stock No. 000664) to develop the colony.

For the Mthfr^em1Adiuj allele (Mthfr*C677T): Plasmids encoding a single guide RNA are designed to introduce a point mutation (GCC to GTC) into the Mthfr gene creating the A262V mutation. Plasmids and cas9 nuclease were introduced into the cytoplasm Stock No. 028709-derived fertilized eggs with well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by sequencing and PCR and further bred to Stock No. 028709 for 2 generations to develop the colony. Sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize oocytes from Stock No. 028709 females.

Expression Data

Expressed Gene	APOE, apolipoprotein E, human
Site of Expression

Control Suggestions

028709 B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J

Additional Information

Considerations for Choosing Controls

Genetics

Trem2^em1Adiuj

Allele Symbol: Trem2^em1Adiuj

Allele Name	endonuclease-mediated mutation 1, MODEL-AD Center
Allele Type	Endonuclease-mediated (Humanized sequence)
Allele Synonym(s)	Jax R27H ki; Trem2 R47H KI
Gene Symbol and Name	Trem2, triggering receptor expressed on myeloid cells 2
Gene Synonym(s)
Strain of Origin	C57BL/6J
Chromosome	17
Molecular Note	The allele was generated by injecting cas9 nuclease and a single guide RNA designed to introduce a R47H point mutation with two silent mutations (lysine AAG>AAA and alanine GCC>GCA) into the gene.

Mthfr^em1Adiuj

Allele Symbol: Mthfr^em1Adiuj

Allele Name	endonuclease-mediated mutation 1, MODEL-AD Center
Allele Type	Endonuclease-mediated (Humanized sequence)
Allele Synonym(s)	Mthfr*A262V
Gene Symbol and Name	Mthfr, methylenetetrahydrofolate reductase
Gene Synonym(s)
Strain of Origin	B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J
Chromosome	4
Molecular Note	CRISPR/Cas9 genome editing is used to introduce a point mutation (GCC to GTC) creating the A262V mutation. This C to T mutation is homologous to the human C677T polymorphism that has been associated with various diseases including Alzheimer's, vascular and metabolic disease.

Apoe^{tm1.1(APOE4)Adiuj}*

Allele Symbol: Apoe^{tm1.1(APOE4)Adiuj}*

Allele Name	targeted mutation 1.1, MODEL-AD Center
Allele Type	Targeted (Inserted expressed sequence, Humanized sequence)
Allele Synonym(s)
Gene Symbol and Name	Apoe, apolipoprotein E
Gene Synonym(s)
Expressed Gene	APOE, apolipoprotein E, human
Strain of Origin	C57BL/6J
Chromosome	7
Molecular Note	The targeting vector contains a 3' FRT flanked neo cassette and 1.5 kb of human APOE gene sequence including exons 2, 3, 4 and a portion of the 3'UTR sequence (the APOE4 isoform) which is used to replace exons 2, 3 and most of exon 4 of the endogenous sequence. The human ApoE sequence carries a 2 nucleotide A to T substitution in intron 3. Flp-mediated recombination removed the FRT-flanked neo cassette.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Research Tools
- Neurobiology Research
- Cardiovascular Research
Neurobiology Research
- Alzheimer's Disease
  - APOE mutants
Cardiovascular Research
- Atherosclerosis
Metabolism Research
- Lipid Metabolism
Mouse/Human Gene Homologs
- Alzheimer's

Mammalian Phenotype Terms by Genotype

References

Additional - Apoe^{tm1.1(APOE4)Adiuj}* related

Additional - Mthfr^em1Adiuj related

Additional - Trem2^em1Adiuj related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- End Point Analysis:Trem2
- Standard PCR:Apoe
- Probe:Apoe-Probe
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, mice heterozygous for Mthfr^em1Adiuj (Mthfr*C677T), and homozygous for Apoe^{tm1.1(APOE*4)Adiuj} (APOE4), App^em1Adiuj (hAbeta) and Trem2^em1Adiuj (Trem2*R47H) may be bred together as they are viable and fertile. Homozygous viability/fertility has not been tested for the Mthfr^em1Adiuj allele [March 2020].
- Additional Breeding and Husbandry Support
Mating System
- See "Breeding Considerations"
Citation
When using the Mthfr*C677T/APOE4/Trem2*R47H mouse strain in a publication, please cite the originating article(s) and include JAX stock #030922 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 or wildtype for Mthfr<em1Adiuj> and Heterozygous for Apoe<tm1.1(APOE*4)Adiuj> and Trem2<em1Adiuj>

Related Products and Services

Frozen Mouse Embryo	B6.Cg-Mthfr<em1Adiuj> Apoe<tm1.1(APOE*4)Adiuj> Trem2<em1Adiu	$2595.00
Frozen Mouse Embryo	B6.Cg-Mthfr<em1Adiuj> Apoe<tm1.1(APOE*4)Adiuj> Trem2<em1Adiu	$2595.00
Frozen Mouse Embryo	B6.Cg-Mthfr<em1Adiuj> Apoe<tm1.1(APOE*4)Adiuj> Trem2<em1Adiu	$3373.50
Frozen Mouse Embryo	B6.Cg-Mthfr<em1Adiuj> Apoe<tm1.1(APOE*4)Adiuj> Trem2<em1Adiu	$3373.50

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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

Terms Of Use

General Terms and Conditions

Questions about Terms of Use

Additional Use Restrictions Apply

The use of this mouse model is subject to the terms and conditions of the Limited License from The Broad Institute.

The use of this mouse model is subject to the terms and conditions of the Limited Use Label License from Caribou Biosciences, Inc.

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

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Also Known As:Mthfr*C677T/APOE4/Trem2*R47H

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Trem2em1Adiuj

Allele Symbol: Trem2em1Adiuj

Mthfrem1Adiuj

Allele Symbol: Mthfrem1Adiuj

Apoetm1.1(APOE*4)Adiuj

Allele Symbol: Apoetm1.1(APOE*4)Adiuj

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 - Apoetm1.1(APOE*4)Adiuj related

Additional - Mthfrem1Adiuj related

Additional - Trem2em1Adiuj related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

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

Live Mouse

Cryorecovery - 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

Also Known As:MthfrC677T/APOE4/Trem2R47H

Trem2^em1Adiuj

Allele Symbol: Trem2^em1Adiuj

Mthfr^em1Adiuj

Allele Symbol: Mthfr^em1Adiuj

Apoe^{tm1.1(APOE4)Adiuj}*

Allele Symbol: Apoe^{tm1.1(APOE4)Adiuj}*

Additional - Apoe^{tm1.1(APOE4)Adiuj}* related

Additional - Mthfr^em1Adiuj related

Additional - Trem2^em1Adiuj related