Overview

Also Known As:Plcg2M28L/APOE4/Trem2R47H

This triple mutant strain carries a humanized ApoE knock-in mutation (sequence coding for isoform E4), CRISPR/Cas9 generated mutant of the Plcg2 gene with the M28L mutation, 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

Plcg2^em3Adiuj

Allele Type	Gene Symbol	Gene Name
Endonuclease-mediated (Humanized sequence)	Plcg2	phospholipase C, gamma 2

View Genetics

Research Applications

Neurobiology Research
Research Tools
Cardiovascular Research
Metabolism Research

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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 mutant of the Plcg2, phospholipase C, gamma 2, gene with the M28L mutation; 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 Trem2 gene 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 homozygous for the Apoe^{tm1.1(APOE*4)Adiuj} and Trem2^em1Adiuj alleles and heterozygous for the Plcg2^em2Adiuj allele are viable and fertile. Homozygous viability/fertility has not been tested for the Plcg2^em2Adiuj allele (June 2018).
As the mice are characterized, we will modify the strain description and add phenotype data.

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 triple mutant line was generated by was generated by utilizing CRISPR/cas9 endonuclease mediated genome editing of the Plcg2 (phospholipase C, gamma 2) gene in
B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J (Stock No. 028709) mice.

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 mice (Stock No. 000664) to develop the colony.

For the Plcg2^em2Adiuj allele:
plasmids encoding a single guide RNA designed to introduce the M28L mutation into the Plcg2 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)Adpmc} Trem2^em1Adpmc/J (Stock No. 028709) to develop the colony.

Expression Data

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

Control Suggestions

B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J mice (Stock No. 028709) may be an appropriate control.

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)
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.

Plcg2^em3Adiuj

Allele Symbol: Plcg2^em3Adiuj

Allele Name	endonuclease-mediated mutation 3, MODEL-AD Center
Allele Type	Endonuclease-mediated (Humanized sequence)
Allele Synonym(s)
Gene Symbol and Name	Plcg2, phospholipase C, gamma 2
Gene Synonym(s)
Strain of Origin	B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J
Chromosome	8
Molecular Note	The allele was generated by injecting cas9 nuclease and a single guide RNA designed to introduce a M28L point mutation.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

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

Mammalian Phenotype Terms by Genotype

References

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

Additional - Plcg2^em3Adiuj related

Additional - Trem2^em1Adiuj related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- End Point Analysis:Trem2
- Standard PCR:Apoe
- End Point Analysis:Plcg2
- 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, and heterozygous for the Plcg2^em2Adiuj allele may be bred. Homozygous viability/fertility has not been tested for the Plcg2^em2Adiuj allele (June 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 for Apoe^{tm1.1(APOE*4)Adiuj} Heterozygous for Plcg2^em3Adiuj Homozygous for Trem2^em1Adiujx Homozygous for Apoe^{tm1.1(APOE*4)Adiuj} Wildtype for Plcg2^em3Adiuj Homozygous for Trem2^em1Adiuj
  ^or
  Homozygous for Apoe^{tm1.1(APOE*4)Adiuj} Wildtype for Plcg2^em3Adiuj Homozygous for Trem2^em1Adiuj X Homozygous for Apoe^{tm1.1(APOE*4)Adiuj} Heterozygous for Plcg2^em3Adiuj Homozygous for Trem2^em1Adiuj
Citation
When using the Plcg2*M28L/APOE4/Trem2*R47H mouse strain in a publication, please cite the originating article(s) and include JAX stock #030674 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(APOE*4)Adiuj> and Trem2<em1Adiuj> Heterozygous or wildtype for Plcg2<em3Adiuj>

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

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Also Known As:Plcg2*M28L/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

Plcg2em3Adiuj

Allele Symbol: Plcg2em3Adiuj

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 - Plcg2em3Adiuj 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

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:Plcg2M28L/APOE4/Trem2R47H

Trem2^em1Adiuj

Allele Symbol: Trem2^em1Adiuj

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

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

Plcg2^em3Adiuj

Allele Symbol: Plcg2^em3Adiuj

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

Additional - Plcg2^em3Adiuj related

Additional - Trem2^em1Adiuj related