B6.Cg-Isr(HSA17)2Mdk App^em1Adiuj/J

Stock number: 038908
Product name: MAPT(H1.0)-GR, hAbeta
Strain synonyms: B6.Cg-Tc(HSA17)2Mdk App^em1Adiuj/J
Research areas: Mouse/Human Gene Homologs, Neurobiology Research

Description

This strain was created by mating B6.Cg-App^em1Adiuj/J (Stock No. 033013) to B6J.B6N-Tc(HSA17)2Mdk/J (Stock No. 035398).

MAPT(H1.0)-GR (Stock No. 035398) mice express the human MAPT (H1 haplotype), MAPT-AS1 transcripts and the typical MAPT protein isoforms. B6J hAbeta (Stock No. 033013) mice carry 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). These mice may be suitable for use in studies related to Alzheimer's disease.

Detailed description

This strain was created by mating B6.Cg-App^em1Adiuj/J (Stock No. 033013) to B6J.B6N-Tc(HSA17)2Mdk/J (Stock No. 035398).

The transchromosomal (Tc) MAPT(H1)-GR (Tc(HSA17)2Mdk) mice contain a syntenic 190 kb region from human chromosome 17 (HSA17) that includes SPPLC2 (signal peptide peptidase 2C) and a modified MAPT (microtubule-associated protein tau), specifically, the H1 MAPT haplotype. The SPPL2C sequence is included because it is embedded entirely in the MAPT-AS1 transcribed region. H1 is linked with late onset Alzheimer's disease (AD) risk, progressive supranuclear palsy, Parkinson's disease, and corticobasal degeneration.

This hAbeta (App^em1Adiuj) mutant strain carries a mutant allele of the App gene containing G601R, F606Y, and R609H point mutations. 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.

Development

This strain was created by mating B6.Cg-App^em1Adiuj/J (Stock No. 033013) to B6J.B6N-Tc(HSA17)2Mdk/J (Stock No. 035398).

The Tc(HSA17)2Mdk allele is created as follows: the mouse Sppl2c (signal peptide peptidase 2C) and Mapt (microtubule-associated protein tau) genes are replaced by human SPPL2C and MAPT (H1 haplotype) genes. The 157 kb deletion on mouse chromosome 11 stretches from, but does not include, Crhr1 to Kansl1, and is replaced by a syntenic 190 kb region from human chromosome 17 (HSA17). The deletion/insertion was made using C57BL/6N-PRX-B6N embryonic stem cells (ES); a loxN-flanked neomycin resistance cassette was removed by transient cre expression prior to establishing the mouse line. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were crossed to C57BL/6J females and maintained on the C57BL/6J background.

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 cas9 nuclease were introduced into the cytoplasm of B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J-derived fertilized eggs with 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 the Apoe and Trem2 alleles were bred out of 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.

Allele

Symbol: App^em1Adiuj
Name: endonuclease-mediated mutation 1, MODEL-AD Center
MGI accession ID: MGI:6164022
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Marker symbol: App
Marker name: amyloid beta (A4) precursor protein
Marker synonyms: AAA; Abeta; ABPP; AD1; Adap; alpha-sAPP; APPI; appican; betaAPP; CTFgamma; Cvap; E030013M08Rik; PN2; PN-II; preA4; protease nexin II
Chromosome: 16
Strain of origin: B6(SJL)-Apoe^{tm1.1(APOE*4)Adiuj} Trem2^em1Adiuj/J
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.

Allele

Symbol: Isr(HSA17)2Mdk
Name: interspecies replacement, human Chr 17, Michael Koob 2
MGI accession ID: MGI:6705045
Generation method: Targeted
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: MAPT(H1.0)-GR
Marker symbol: Isr(HSA17)2Mdk
Marker name: interspecies replacement, human Chr 17, Michael Koob 2
Marker synonyms: Tc(HSA17)2Mdk
Chromosome: 11
Strain of origin: C57BL/6NTac
Molecular note: A 157 kb deletion on mouse chromosome 11 stretching from, but not including, Crhr1 to Kansl1 is replaced by a syntenic 190 kb region from human chromosome 17 (HSA17). The mouse Sppl2c (signal peptide peptidase 2C) and Mapt (microtubule-associated protein tau) genes are replaced by human SPPL2C and MAPT (H1 haplotype) genes. A loxN-flanked neomycin resistance cassette was removed by transient cre expression.