NOD.Cg-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas Prkdc^scid Il2rg^tm1Wjl/SentJ

Stock number: 041591
Product name: 5xFAD-NSG
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Neurobiology Research, Research Tools, Virology Research

Description

5xFAD-NSG mice overexpress both mutant human amyloid beta precursor protein (APP) with the Swedish (K670N, M671L), Florida (I716V), and London (V717I) Familial Alzheimer's Disease (FAD) mutations and human presenilin 1 (PSEN1), harboring two FAD mutations, M146L and L286V, on the immunodeficient NSG background. These mice may be useful when studying Alzheimer's disease and evaluating relevant human cell-based therapeutics.

Detailed description

5xFAD-NSG mice are NOD.scid.Il2Rg^null ("NSG"; Stock No. 005557) animals also harboring 5xFAD transgenes (MMRRC Strain No. 034848-JAX).

NSG mice are immunodeficient; they have no mature T cells or B cells, lack functional natural killer (NK) cells, have reduced numbers of lymphocytes and myeloid dendritic cells, and are deficient in cytokine signaling. The NSG platform is permissive for xenograft/human tumor growth and for evaluating human cell-based therapeutics. The 5xFAD transgenes, mutant human amyloid beta precursor protein (APP) with the Swedish (K670N, M671L), Florida (I716V), and London (V717I) Familial Alzheimer's Disease (FAD) mutations and human presenilin 1 (PSEN1), harboring two FAD mutations M146L and L286V, are expressed under direction of the mouse Thy1 promoter to drive overexpression in the brain. The 5xFAD transgenic founder line 6799 was determined to have high APP expression correlating with high burden and accelerated accumulation of the 42 amino acid species of beta-amyloid (Aβ-42).

Mice that are hemizygous for the 5xFAD transgenes [Tg(APPSwFlLon, PSEN1*M146L*L286V)6799Vas], homozygous for the Prkdc^scid allele, and homozygous (or hemizygous) for the X-linked Il2rg^tm1Wjl knock-out allele are viable and fertile. Mice hemizygous for the 5xFAD transgene develop Aβ-42 plaques in the brain, starting ~13 weeks of age, as well as microgliosis, neuroinflammation, and behavior changes that progress with age. Due to the findings that parental origin of the 5xFAD transgene can affect plaque deposition (Sasmita et al., 2025 Neuron [PMID:39837326]), it is recommended this line be maintained by mating females that are non-carrier (WT) for 5xFAD transgenes, homozygous for the Prkdc^scid allele, and homozygous for the X-linked Il2rg^tm1Wjl knock-out allele with males that are hemizygous for the 5xFAD transgenes, homozygous for the Prkdc^scid allele and hemizygous for the X-linked Il2rg^tm1Wjl knock-out allele.

5xFAD-NSG mice may be useful when studying Alzheimer's disease and for evaluating relevant human cell-based therapeutics.

These mice are severely immunocompromised. SPF (specific pathogen-free) husbandry conditions are recommended. Please see our NSG housing and breeding information for additional details.

Development

The 5xFAD-NSG mouse line was generated by backcrossing hemizygous 5xFAD transgenic mice (MMRRC Strain No. 034848-JAX) on to an NSG (Stock No. 005557) background for at least 8 generations. Additional details about the alleles expressed in this mouse line are listed below.

The 5xFAD transgenic transgenic line was generated as follows: A transgene that was designed with a mutant human amyloid beta (A4) precursor protein (APP) cDNA sequence (altered to include the APP K670N/M671L (Swedish) + I716V (Florida) + V717I (London) Familial Alzheimer's Disease (FAD) mutations) inserted into exon 2 of the mouse Thy1 gene. A second transgene was designed with a mutant human presenilin 1 (Alzheimer disease 3) (PSEN1 or PS1) cDNA sequence (altered to include the PS1 M146L + L286V FAD mutations) inserted into exon 2 of the mouse Thy1 gene. Both transgenes were added together in equal proportions and co-injected into the pronuclei of single-cell "C57BL/6XSJL" hybrid embryos. Founders from the highest APP expressing line (Tg6799) were bred with (B6SJL)F1 mice for more than 10 generations with stable germ-line transmission and expression of both transgenes, demonstrating that these "5XFAD" mice breed as single transgenics. The mice were then backcrossed to C57BL/6J mice using a speed congenic protocol and the retinal degeneration allele Pde6b^rd1 was bred out of the strain. Of note, the APP transgene includes the 5' untranslated region and thus contains a putative interleukin-1beta translational enhancer element. The 5xFAD transgenes are reported to have co-inserted onto Chromosome 3 ~63 Mb [MGI:3693208].

NSG mice are non-obese diabetic animals (NOD/ShiLtJ) harboring the spontaneous severe combined immunodeficiency mutation (Prkdc^scid) on chromosome 16 and the Il2Rg^null mutation at ~101 Mbp on the X chromosome (Il2rg^tm1Wjl created by Dr. Warren J. Leonard [NIH]).

Dr. Charles Sentman acquired some hemizygous B6-congenic 5xFAD transgenic mice (MMRRC Strain No. 034848-JAX), along with some NSG mice, and continuously backcrossed the 5xFAD transgene on to the NSG background for at least 8 generations. Donor males that were hemizygous for Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas, homozygous for Prkdc^scid and hemizygous for the X-linked Il2rg^tm1Wjl alleles were provided to The Jackson Laboratory. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize NSG oocytes (Stock No. 005557).

Allele

Symbol: Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas
Name: transgene insertion 6799, Robert Vassar
MGI accession ID: MGI:3693208
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: 5XFAD; 5XFAD APP/PS1; 5XFAD line Tg6799; Tg(APP*Swe*Fl*Lon,PSEN1*M146L*L286V)6799Vas; Tg-5xFAD; Tg6799
Marker symbol: Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas
Marker name: transgene insertion 6799, Robert Vassar
Marker synonyms: 5XFAD; 5XFAD line Tg6799; Tg6799
Chromosome: 3
Strain of origin: (C57BL/6 x SJL)F1
Expressed genes: PSEN1,presenilin 1,human; APP695,amyloid beta (A4) precursor protein (chimeric),mouse/human chimera
Site of expression: Expression is directed to the brain by neural-specific elements of the mouse Thy1 promoter

Molecular note: Four familial Alzheimer disease- (FAD-) associated mutations were introduced into a single human amyloid precursor protein cDNA: the "Swedish" double mutation (K670N/M671L); the "Florida" mutation (I716V); and the "London" mutation (V717I). Two FAD-associated mutations, M146L and L286V, likewise were introduced into a human presenilin 1 cDNA. Each cDNA was then cloned independently into the mouse thymus cell antigen 1 gene, replacing a segment that contains thymus-specific elements so that expression of the transgenes is targeted only to the brain. Equal molar amounts of the two transgenes were coinjected into pronuclei of single-celled embryos.
General note: Three transgenic lines coexpressing the APP and PSEN1 proteins at high, medium and low levels, respectively designated Tg6799, Tg7031, and Tg7092, were propagated for analysis, most of which employed Tg6799.

Phenotypic Similarity to Human Syndrome: Macular Degeneration, Age-Related J:214858.

Allele

Symbol: Prkdc^scid
Name: severe combined immunodeficiency
MGI accession ID: MGI:1857113
Generation method: Spontaneous
Synonyms: scid
Marker symbol: Prkdc
Marker name: protein kinase, DNA activated, catalytic polypeptide
Marker synonyms: DNAPDcs; DNAPK; DNA-PK; DNAPKc; DNA-PKC; DNA-PKcs; DNPK1; DOXNPH; dxnph; HYRC; HYRC1; IMD26; p350; S473K; slip; XRCC7
Chromosome: 16
Strain of origin: C.BKa-Igh^b/Icr
Site of expression: T and B lymphocytes.
Molecular note: A T-to-A transversion point mutation at a position corresponding to codon 4046 (codon 4095 in transcript ENSMUST00000023352.8) created a premature stop codon (p.Y4046*).

Allele

Symbol: Il2rg^tm1Wjl
Name: targeted mutation 1, Warren J Leonard
MGI accession ID: MGI:1857455
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: [KO]gammac; CD132^-; gammac^-; gc^-; Il2rg^tm1Wjll; IL2Rgamma^null
Marker symbol: Il2rg
Marker name: interleukin 2 receptor, gamma chain
Marker synonyms: [g]c; Ab2-183; Cd132; CIDX; common cytokine receptor gamma chain; common gamma chain; gamma C receptor; gamma(c); gammaC; gc; IL-2RG; IMD4; P64; SCIDX; SCIDX1
Chromosome: X
Strain of origin: 129S4/SvJae
Site of expression: Primarily lymphoid cells.
Molecular note: A neomycin resistance cassette replaced part of exon 3 and all of exons 4 - 8 of the gene, resulting in the loss of most of the extracellular domain and all of the transmembrane and cytoplasmic domains of the protein.