Overview

Also Known As:Il1rap^ex3KO/hAbeta/APOE4/Trem2*R47H

This model has a CRISPR/cas9-generated deletion of exon 3 of the Il1rap (interleukin 1 receptor accessory protein) gene. The IL1RAP intronic variant rs12053868 is associated with amyloid accumulation in the ADNI cohort.
This mutation is maintained with the hAbeta/APOE4/Trem2*R47H mutations (Stock No. 030670) intended to increase risk of late-onset Alzheimer's disease: a humanized Apoe knock-in mutation (sequence coding for human isoform E4), a CRISPR/Cas9-generated App allele with a humanized Abeta1-42 region (G601R, F606Y, R609H in the mouse gene, corresponding to amino acid positions 676, 681, 684 in the human APP locus) and a CRISPR/Cas9-generated R47H point mutation of the Trem2 gene.

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

App^em1Adiuj

Allele Type	Gene Symbol	Gene Name
Endonuclease-mediated (Humanized sequence)	App	amyloid beta (A4) precursor protein

Il1rap^em2Adiuj

Allele Type	Gene Symbol	Gene Name
Endonuclease-mediated (Null/Knockout)	Il1rap	interleukin 1 receptor accessory protein

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

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

View Genetics

Research Applications

Neurobiology Research
Mouse/Human Gene Homologs

View All Research Applications

Details

Detailed Description

Il1rap^ex3KO/hAbeta/APOE4/Trem2*R47H quadruple mutant strain carries a mutant allele of the Il1rap lacking exon 3 (modeling human intronic variant rs12053868), 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 mutant allele of the App gene containing G601R, F606Y and R609H point mutations and a knock-in of a point mutation into mouse Trem2 gene containing an R47H point mutation with two silent mutations.

The targeted Il1rap gene encodes a component of the interleukin 1 receptor complex, which initiates signaling events that result in the activation of interleukin 1-responsive genes. The encoded gene has also been associated with amyloid accumulation in the Alzheimer’s Disease Neuroimaging Initiative (ADNI) cohort. 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 App gene encodes amyloid beta precursor protein, a transmembrane cell surface receptor that is cleaved by secretases. Mutations in this gene have been associated with Alzheimer's disease. 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 homozygous for Il1rap^em2Adiuj (Il1rap^ex3KO), Apoe^{tm1.1(APOE*4)Adiuj} (APOE4), App^em1Adiuj (hAbeta) and Trem2^em1Adiuj (Trem2*R47H) are viable and fertile [July 2020]. If additional characterization of these Il1rap^ex3KO/hAbeta/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 Il1rap^ex3KO/hAbeta/APOE4/Trem2*R47H quadruple mutant line was generated by utilizing CRISPR/Cas9 endonuclease-mediated genome editing of the Il1rap gene in C57BL/6J-congenic Apoe^{tm1.1(APOE*4)Adiuj} App^em1Adiuj Trem2^em1Adiuj mice (available as Stock No. 030670).
Stock No. 030670 was generated by utilizing CRISPR/Cas9 endonuclease-mediated genome editing of the App gene in C57BL/6J-congenic Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj mice (available as Stock No. 028709).

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 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 App^em1Adiuj allele (hAbeta): Plasmids encoding a single guide RNA designed to introduce the G601R, F606Y, and R609H point mutations into the App gene and the cas9 nuclease were introduced into the cytoplasm of Stock No. 028709-derived fertilized eggs with well recognized pronuclei. G601R corresponds to a GGA to CGA nucleotide change, F606Y represents a TTT to TAT nucleotide mutation, and R609H incorporates a CGC to CAT nucleotide change. Correctly targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by sequencing and PCR and further bred to Stock No. 028709 to develop the colony.
Important note about App isoforms and exon numbering: The App-201 isoform (695 aa protein) is encoded by 16 exons, of which the Abeta sequence is encoded by exon 14. The App-206 isoform (770 aa protein) is encoded by 18 exons, of which the Abeta sequence is encoded by exon 16.

For the Il1rap^em2Adiuj allele (Il1rap^ex3KO): Plasmids encoding a single guide RNA designed to delete exon 3 of the Il1rap locus and the cas9 nuclease were introduced into the cytoplasm of Stock No. 030670-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. 030670 to develop the colony.

Expression Data

Expressed Gene	APOE, apolipoprotein E, human
Site of Expression

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.

App^em1Adiuj

Allele Symbol: App^em1Adiuj

Allele Name	endonuclease-mediated mutation 1, MODEL-AD Center
Allele Type	Endonuclease-mediated (Humanized sequence)
Allele Synonym(s)
Gene Symbol and Name	App, amyloid beta (A4) precursor protein
Gene Synonym(s)
Strain of Origin	B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J
Chromosome	16
Molecular Note	CRISPR/Cas9 genome editing methodologies are used to introduce G601R, F606Y, and R609H point mutations into the App gene. G601R corresponds to a GGA to CGA nucleotide change, F606Y represents a TTT to TAT change, and R609H incorporates a CGC to CAT nucleotide change.

Il1rap^em2Adiuj

Allele Symbol: Il1rap^em2Adiuj

Allele Name	endonuclease-mediated mutation 2, MODEL-AD Center
Allele Type	Endonuclease-mediated (Null/Knockout)
Allele Synonym(s)	Il1rap^ex3KO
Gene Symbol and Name	Il1rap, interleukin 1 receptor accessory protein
Gene Synonym(s)
Strain of Origin	B6.Cg-Apoe^{tm1.1(APOE*4)Adiuj} App^em1Adiuj Trem2^em1Adiuj/J
Chromosome	16
Molecular Note	CRISPR/Cas9 genome editing is used to delete exon 3.

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:

Neurobiology Research
- Alzheimer's Disease
  - APOE mutants
  - APP mutants
Mouse/Human Gene Homologs
- Alzheimer's

Mammalian Phenotype Terms by Genotype

References

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

Additional - App^em1Adiuj related

Additional - Il1rap^em2Adiuj related

Additional - Trem2^em1Adiuj related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, mice homozygous for Apoe^{tm1.1(APOE*4)Adiuj} (APOE4), Il1rap^em2Adiuj (Il1rap^ex3KO), App^em1Adiuj (hAbeta) and Trem2^em1Adiuj (Trem2*R47H) may be bred together as they are viable and fertile. [July 2020]
- Additional Breeding and Husbandry Support
Citation
When using the Il1rap^ex3KO/hAbeta/APOE4/Trem2*R47H mouse strain in a publication, please cite the originating article(s) and include JAX stock #032777 in your Materials and Methods section.

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

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Also Known As:Il1rapex3KO/hAbeta/APOE4/Trem2*R47H

Donating Investigator

Genetic overview

Research Applications

Detailed Description

Development

Expression Data

Trem2em1Adiuj

Allele Symbol: Trem2em1Adiuj

Appem1Adiuj

Allele Symbol: Appem1Adiuj

Il1rapem2Adiuj

Allele Symbol: Il1rapem2Adiuj

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 - Appem1Adiuj related

Additional - Il1rapem2Adiuj related

Additional - Trem2em1Adiuj related

Genotyping Protocols

Breeding Considerations

Citation

Strain Interest Registration

Contact Information

Interest

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:Il1rap^ex3KO/hAbeta/APOE4/Trem2*R47H

Trem2^em1Adiuj

Allele Symbol: Trem2^em1Adiuj

App^em1Adiuj

Allele Symbol: App^em1Adiuj

Il1rap^em2Adiuj

Allele Symbol: Il1rap^em2Adiuj

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

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

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

Additional - App^em1Adiuj related

Additional - Il1rap^em2Adiuj related

Additional - Trem2^em1Adiuj related