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5×FAD five-transgenic mice

Jiangsu Aniphe BioLab boasts a mature 5×FAD transgenic mouse model platform, supported by standardized breeding management, precise genotyping, and a multi‑dimensional phenotypic validation system, ensuring the models’ stability and reliability. The company offers models spanning diverse stages, from early Aβ deposition to late‑stage cognitive impairment, thereby meeting research needs such as Alzheimer’s disease mechanism studies, candidate drug screening, and efficacy evaluation, and providing high‑quality animal models to advance the development of therapeutics for neurodegenerative diseases.

Strain Introduction

The 5×FAD (5-Familial Alzheimer’s Disease mutation) mouse is a transgenic model that rapidly and reproducibly recapitulates the Aβ cascade–driven pathology of Alzheimer’s disease (AD). This model simultaneously overexpresses three pathogenic mutations in the human APP gene and two pathogenic mutations in the human PSEN1 gene—five familial AD mutations in total—under the control of the Thy1 neuron‑specific promoter, resulting in high‑level expression throughout the central nervous system.

In 5×FAD mice, intracellular Aβ accumulation emerges around 1.5 months after birth, while extracellular amyloid plaque deposition becomes apparent by approximately 2 months. By 6 months, plaque burden is already extensive, accompanied by marked microglial and astrocytic activation, synaptic loss, and robust deficits in spatial learning and memory. Due to its early onset of pathological features, rapid progression, and heavy plaque load, the 5×FAD model has become one of the most widely used preclinical platforms for studying Aβ‑driven Alzheimer’s disease worldwide.

Product Advantages

Pathological progression is rapid, shortening the experimental timeline.

At 1.5 months of age, Aβ accumulates within neurons; by 2 months, extracellular amyloid plaques become apparent; and pharmacological efficacy can be evaluated at 3–4 months, significantly shortening the traditional model’s 9–12‑month experimental timeline.

Five pathogenic mutations, with high clinical and pathological similarity.

Carrying five familial Alzheimer’s disease–associated clinical mutations, including APP and PSEN1, it efficiently recapitulates Aβ overaccumulation and is well suited for studying Aβ metabolism and the mechanisms of secretory enzymes.

The phenotype is homogeneous and stable, with excellent experimental reproducibility.

Mutant‑linked integration ensures stable inheritance through hemizygous breeding; minimal inter‑individual patch variability makes it well suited for high‑throughput drug screening.

 

Data parameters

Strain Name B6.Cg-Tg(APPSwFlLon,PSEN1M146LL286V)6799Vas/Mmjax
Targeted gene APP, PSEN1
Mutation site APP K670_M671delinsNL (Swedish), APP I716V (Florida), APP V717I (London), PSEN1 M146L (A>C), PSEN1 L286V
Types of gene editing APP-transgenic; PSEN1-transgenic
Genetic background C57BL/6
Microbial Grade SPF grade

Validate data

Early-onset Aβ plaque–rich classic AD model

5xFAD mice express human APP and PSEN1 transgenes, carrying five AD‑associated mutations: the Swedish (K670N/M671L), Florida (I716V), and London (V717I) familial Alzheimer’s disease (FAD) mutations in APP, as well as two FAD mutations—M146L and L286V—in human PS1. These transgenes are driven by the Thy1 promoter, which mediates brain‑specific expression under the control of mouse neuronal regulatory elements.

 

 

Image source: [2]

At 1.5 months of age, 5×FAD mice already exhibit abundant β‑amyloid (Aβ) in the brain, and neuritic plaques (NP) begin to appear by 2 months of age. The pathological phenotype of 5×FAD mice includes amyloid plaque accumulation, neuronal loss, glial cell activation, and cognitive deficits. These 5XFAD transgenic mice rapidly recapitulate the key hallmarks of amyloid pathology in Alzheimer’s disease and may serve as a valuable model for studying Aβ‑42‑induced neurodegeneration and amyloid plaque formation within neurons.

 

 

Brain volume increases with age.

The brain volume of 5xFAD mice increases with age, and both male and female brains exhibit significant genotype‑dependent differences by 8 months of age in females.

 

 

Sources of brain volume across different sexes and age groups [3]

 

Amyloid deposition and glial proliferation

Amyloid deposition in 5xFAD mice begins in the deep and lower cortical layers at 2 months of age and, with advancing age, spreads to other brain regions. In the B6SJL hybrid background, the amyloid phenotype in 5xFAD mice is more stable.

 

 

Amyloid deposition and glial activation in the 5XFAD model; image source: [2]

 

Astrocytic and microglial proliferation begin to emerge around two months, progressing in parallel with amyloid plaque deposition.

 

 

Rapid cerebral amyloid deposition and glial activation in 5XFAD Tg6799 mice at 2, 4, 6, and 9 months of age; image source: [2].

 

Neuronal loss

An upward trend was observed in 5xFAD mice at 3 months of age, with neuronal loss in the cortical V layer and deeper layers.

 

 

Figure 5 from Tg6799 shows loss of cortical layer V neurons and lower-layer neurons; the image is reproduced from [2].

 

Synaptic loss

The synaptic markers synaptophysin, syntaxin, and PSD-95 exhibit decreased expression with advancing age in mice.

 

 

Image source: [3]

 

Changes in LTP/LTD

Basal synaptic transmission and LTP in the CA1 region of the hippocampus begin to deteriorate between 4 and 6 months of age, yet paired-pulse facilitation remains normal at least until 6 months. Deficits in basal synaptic transmission were also observed in layer V of the somatosensory cortex of 6-month-old 5xFAD mice. At approximately one month and again between 11 and 15 months, there were no differences between transgenic and wild-type mice in the amplitude or frequency of spontaneous excitatory postsynaptic currents in the radiatum layer of the hippocampus.

 

 

Age-dependent impairment of LTP in hippocampal slices from 5XFAD mice, but not paired-pulse facilitation; image reproduced from [4].

 

Mouse behavioral phenotype

At 4–5 months of age, 5XFAD mice exhibit impaired learning and memory, as evidenced by reduced spontaneous alternation performance in the Y-maze. They also show diminished learning and memory abilities in cued and contextual fear‑conditioning tasks. Furthermore, deficits in spatial learning and memory are observed in the water maze task and other hippocampus‑dependent memory paradigms.

 

 

Memory deficits in 5XFAD mice increase with age; image source: [2]

Application areas

● Research on the pathogenesis of AD: By investigating the pathological features and behavioral alterations in 5xFAD mice, it is possible to gain deeper insights into the pathogenic mechanisms of Alzheimer’s disease, particularly the pathological processes associated with β‑amyloid deposition and synaptic dysfunction.
● Drug screening and therapeutic evaluation: 5xFAD mice can be used to evaluate the efficacy of potential therapies and drugs for Alzheimer’s disease, including anti-amyloid agents and neuroprotective compounds.
● Research on Genetic and Environmental Factors: By conducting comparative studies with other transgenic mouse models, it is possible to elucidate the interactions between genetic and environmental factors in the pathogenesis of Alzheimer’s disease.

Breeding Techniques and Precautions

Strictly control inbreeding systems to mitigate genetic diversity.

Genotype Confirmation and Marker Management

Prior to the experiment, PCR-based genotyping must be performed to accurately distinguish positive mice from negative control mice, and information such as ear tags, dates of birth, and sex should be meticulously recorded to prevent errors in experimental grouping.

 

Strictly control environmental stimuli to reduce experimental stress.

Maintain a stable rearing environment.

Maintain constant temperature and humidity, a 12-hour light–dark cycle, and minimize environmental stressors such as noise and frequent cage changes to reduce the impact of stress on neuroinflammation and behavioral outcomes.

Standardize breeding management to improve the survival rate of pups.

Reasonable grouping and housing in cages

It is recommended to house mice of the same sex and age in groups (3–5 per cage) to prevent overcrowding; older male mice should be monitored for aggressive behavior, and housing density should be adjusted promptly when necessary.

Monitor age-related spontaneous lesions and eliminate experimental confounding.

Daily health monitoring

Regularly monitor the mice’s mental state, activity level, coat condition, and body weight; during the 5×FAD aging phase, reduced activity and declining physical condition may be observed.

Control male mouse fighting and standardize breeding practices.

Diet and Fluid Management

Maintain consistent feed and water supply, and avoid frequent changes in feed brands or housing conditions; if a high-fat diet is used as an intervention, include appropriate control groups and record body weight changes.

Monitor age-related spontaneous lesions and eliminate experimental confounding.

Reduce experimental stress

Procedures such as handling, cage changes, and drug administration should be performed gently and according to a standardized protocol to minimize stress‑induced effects on inflammatory markers, behavioral outcomes, and experimental results.

Literature support

Scientific publications based on this model, as cited by Aniphe customers, are supported for submission to SCI-indexed journals. (Original paper link)
Int J Mol Sci.  2024 Feb 14;25(4):2293.

Frequently Asked Questions

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Q1: What are the key differences between 5×FAD and APP/PS1, as well as 3xTg-AD? Which one is better to choose?

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A: 5×FAD: Plaque formation is fastest (at 2 months), making it suitable for rapid screening of Aβ‑related drugs, anti‑amyloid therapies, immunotherapies, and BACE1/γ‑secretase inhibitors. No mature tau tangles are present, and cerebral amyloid angiopathy is mild.
APP/PS1: Plaques develop between months 6 and 9, with CAA being more pronounced, making it suitable for vascular pathology studies.
3xTg-AD: Exhibits both Aβ and tau pathologies, yet progresses very slowly (plaque accumulation >12 months), making it well-suited for long-term studies of tau inhibitors.
Conclusion: For speed, choose the 5×FAD model; for vascular pathology, select APP/PS1; and for tau tangles, opt for 3xTg or a hybrid model.

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Q2: What age should be chosen for the 5×FAD mouse model in experiments?

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A: The appropriate age should be selected based on the research objectives: for studies on Aβ production and deposition, 2–4-month-old animals are suitable; for investigations of inflammation, synaptic damage, and cognitive function, animals aged 4–6 months or older are typically chosen.

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Q3: What gender-related differences does this model exhibit?

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A: Pathological progression is markedly faster in female mice than in males. Female 5×FAD mice exhibit earlier and more severe brain Aβ deposition, greater glial activation, and the onset of cognitive deficits compared with age-matched male mice. This phenomenon is often attributed to estrogen-mediated regulation of the Thy-1 promoter (which drives transgene expression) or sex-specific metabolic differences. Therefore, when designing experiments, it is advisable to treat sex as an independent variable or to use only one sex throughout.

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