Overview

Also Known As:Clasp2L163P/APOE4/Trem2R47H

This triple mutant strain carries a humanized ApoE knock-in mutation (sequence coding for isoform E4), a CRISPR/cas9-generated L163P point mutation of the Clasp2 (CLIP associating protein 2) gene 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

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

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

Clasp2^em1Adiuj

Allele Type	Gene Symbol	Gene Name
Endonuclease-mediated (Not Specified)	Clasp2	CLIP associating protein 2

View Genetics

Research Applications

Cardiovascular Research
Research Tools
Metabolism Research
Neurobiology Research

View All Research Applications

Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

This triple mutant strain carries 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); a CRISPR/cas9-generated L163P point mutation of the Clasp2 (CLIP associating protein 2) and a knock-in of a point mutation into mouse Trem2, triggering receptor expressed on myeloid cells 2, gene containing a R47H point mutation, with two silent mutations. 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 Clasp2 gene encodes a microtubule plus-end tracking protein. The targeted Trem2 gene encodes a protein that is part of a receptor signaling complex with TYRO protein tyrosine kinase binding protein, which activates macrophages and dendritic cells during immune responses. Loss-of-function variants of the TREM2 gene are associated with Alzheimer's Disease. The R47H point mutation has been shown to increase Alzheimer's disease risk. Mice that are homozygous for the Apoe^{tm1.1(APOE*4)Adiuj} and Trem2^em1Adiuj alleles, and heterozygous for the Clasp2^em1Adiuj allele are viable and fertile. Homozygous viability/fertility has not been tested for the Clasp2^em1Adiuj allele (March 2018). As the mice are characterized, we will modify the strain description and add phenotype data.

Development

This triple mutant line was generated by utilizing CRISPR/cas9 endonuclease mediated genome editing of the Clasp2 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 (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 (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 of 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 Clasp2^em1^Adiuj allele: plasmids encoding a single guide RNA designed to introduce the L163P point mutation (CTG>CCA) into the Clasp2 gene and the cas9 nuclease were introduced into the cytoplasm of B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J (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 B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J (Stock No. 028709). Sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J oocytes (Stock No. 028709).

Expression Data

Expressed Gene	APOE, apolipoprotein E, human
Site of Expression
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.

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)	APOE^epsilon4
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.

Clasp2^em1Adiuj

Allele Symbol: Clasp2^em1Adiuj

Allele Name	endonuclease-mediated mutation 1, MODEL-AD Center
Allele Type	Endonuclease-mediated (Not Specified)
Allele Synonym(s)
Gene Symbol and Name	Clasp2, CLIP associating protein 2
Gene Synonym(s)
Strain of Origin	B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J
Chromosome	9
Molecular Note	A plasmids encoding a single guide RNA designed to introduce the L163P point mutation (CTG to CCA) into the gene and the cas9 nuclease were introduced into the cytoplasm of fertilized eggs with well recognized pronuclei.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Cardiovascular Research
Research Tools
- Cardiovascular Research
- Neurobiology Research
Metabolism Research
- Lipid Metabolism
Neurobiology Research
- Alzheimer's Disease
  - APOE mutants

Mammalian Phenotype Terms by Genotype

References

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

Additional - Clasp2^em1Adiuj related

Additional - Trem2^em1Adiuj related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- End Point Analysis:Trem2
- Standard PCR:Apoe
- Standard PCR:Apoe
- End Point Analysis:Clasp2
- Probe:Apoe-Probe
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, mice homozygous for the Apoe^{tm1.1(APOE*4)Adiuj} and Trem2^em1Adiuj alleles, and heterozygous for the Clasp2^em1Adiuj allele may be bred.
Homozygous viability/fertility has not been tested for the Clasp2^em1Adiuj allele (March 2018).
As the mice are characterized, we will modify the strain description if necessary and add data related to viability and fertility.
- Additional Breeding and Husbandry Support
Mating System
- Homozygous Apoe, homozygous Trem2 x Homozygous Apoe, homozygous Trem2
Citation
When using the Clasp2*L163P/APOE4/Trem2*R47H mouse strain in a publication, please cite the originating article(s) and include JAX stock #031944 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
	Homozygous for Apoe<tm1.1(APOE4)Adiuj>, heterozygous for Clasp2<em1Adiuj>, and homozygous for Trem2<em1Adiuj> or homozygous for Apoe<tm1.1(APOE4)Adiuj>, wildtype for Clasp2<em1Adiuj>, and homozygous for Trem2<em1Adiuj>

Related Products and Services

Frozen Mouse Embryo	B6.Cg-Apoe<tm1.1(APOE*4)Adiuj> Clasp2<em1Adiuj> Trem2<em1Adi	$2595.00
Frozen Mouse Embryo	B6.Cg-Apoe<tm1.1(APOE*4)Adiuj> Clasp2<em1Adiuj> Trem2<em1Adi	$2595.00
Frozen Mouse Embryo	B6.Cg-Apoe<tm1.1(APOE*4)Adiuj> Clasp2<em1Adiuj> Trem2<em1Adi	$3373.50
Frozen Mouse Embryo	B6.Cg-Apoe<tm1.1(APOE*4)Adiuj> Clasp2<em1Adiuj> Trem2<em1Adi	$3373.50

Payment Terms and Conditions

Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.

The Jackson Laboratory's Genotype Promise

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

Also Known As:Clasp2*L163P/APOE4/Trem2*R47H

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Trem2em1Adiuj

Allele Symbol: Trem2em1Adiuj

Apoetm1.1(APOE*4)Adiuj

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

Clasp2em1Adiuj

Allele Symbol: Clasp2em1Adiuj

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 - Clasp2em1Adiuj 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

Also Known As:Clasp2L163P/APOE4/Trem2R47H

Trem2^em1Adiuj

Allele Symbol: Trem2^em1Adiuj

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

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

Clasp2^em1Adiuj

Allele Symbol: Clasp2^em1Adiuj

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

Additional - Clasp2^em1Adiuj related

Additional - Trem2^em1Adiuj related