Name: B6-hATXN3 Mouse
Brand: サイヤジェン（Cyagen）
SKU: C001398

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B6-hATXN3 Mouse

製品名

B6-hATXN3 Mouse

製品ID

C001398

系統名

C57BL/6NCya-Atxn3^tm1(hATXN3)/Cya

背景情報

C57BL/6NCya

Note

One of Cyagen's HUGO-GT^® (Humanized Genomic Ortholog for Gene Therapy) Mouse Strains

状況

このマウス系統を論文で使用する場合は、「B6-hATXN3 Mouse（カタログ番号C001398）はサイアジェンから購入しました。」と引用してください。

HUGO-GT Humanized Models

Neurodegenerative Diseases

製品タイプ

年齢

遺伝子型

性別

数量

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HUGO-GT Humanized Models

Neurodegenerative Diseases

基本情報

検証 Data

系統詳細

Spinocerebellar ataxias (SCAs) are a group of genetic diseases that mainly manifest as chronic progressive ataxia, such as limping, sudden falls, and difficulty in pronunciation. The main lesion sites of these diseases are the cerebellum and its associated tissues. They are mostly inherited in an autosomal dominant manner, but there are also autosomal recessive and X-linked inheritance types. The average incidence of SCA is 2.7 per 100,000 people ^[1]. SCA can be divided into repeat expansion type and non-repeat expansion type according to the genetic mutation type. Among them, repeat expansion type includes polyglutamine SCA and non-translated region repeat expansion type SCA. Spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph disease (MJD), belongs to polyglutamine SCA and is the most common dominant hereditary ataxia. The pathogenesis of SCA3 is the loss of neurotransmitters caused by CAG repeat expansion in the ATXN3 gene. This expansion results in a long polyglutamine (polyQ) domain in the Ataxin 3 protein, leading to protein aggregation and dysfunction of the ubiquitin-proteasome system. The CAG repeat number in the healthy human ATXN3 gene ranges from 12 to 44, while the polyQ domain of SCA3 patients abnormally increases, with CAG repeat numbers ranging from 56 to 87. Individuals with CAG repeat numbers between 45 and 55 exhibit incomplete penetrance of SCA3 symptoms. Like other PolyQ diseases, the CAG repeat number is negatively correlated with the age of onset of SCA3 and positively correlated with the severity of the disease ^[2-3].

Currently, most SCA treatments targeting the ATXN3 gene are in the early stages of development and mainly involve reducing abnormal ATXN3 expression through means such as miRNA or ASO drugs. The Ataxin 3 protein in mice does not contain or only contains a shorter polyQ structure. Considering the differences between humans and mice in terms of genes, humanizing mouse genes can help accelerate these treatments into clinical stages. This strain is a mouse Atxn3 gene humanized model that can be used for research on Spinocerebellar ataxia type 3 (SCA3) ^[4-9]. The homozygous B6-hATXN3 mice are viable and fertile. Additionally, based on the independently developed TurboKnockout fusion BAC recombination technology, Cyagen can also generate hot mutation models based on this strain and provide customized services for specific mutations to meet experimental needs in pharmacology.

参考文献

Ruano L, Melo C, Silva MC, Coutinho P. The global epidemiology of hereditary ataxia and spastic paraplegia: a systematic review of prevalence studies. Neuroepidemiology. 2014;42(3):174-83.

Paulson H. Machado-Joseph disease/spinocerebellar ataxia type 3. Handb Clin Neurol. 2012;103:437-49.

McLoughlin HS, Moore LR, Paulson HL. Pathogenesis of SCA3 and implications for other polyglutamine diseases. Neurobiol Dis. 2020 Feb;134:104635.

Cemal CK, Carroll CJ, Lawrence L, Lowrie MB, Ruddle P, Al-Mahdawi S, King RH, Pook MA, Huxley C, Chamberlain S. YAC transgenic mice carrying pathological alleles of the MJD1 locus exhibit a mild and slowly progressive cerebellar deficit. Hum Mol Genet. 2002 May 1;11(9):1075-94.

Mcloughlin H S , Moore L R , Paulson H L .Pathogenesis of SCA3 and implications for other polyglutamine diseases[J].Neurobiol Dis, 2020.

Zhe,Long,Zhao,et al.Two novel SNPs in ATXN3 3' UTR may decrease age at onset of SCA3/MJD in Chinese patients.[J].Plos One, 2015.

Krauss S , Nalavade R , Weber S ,et al.Upregulation of miR-25 and miR-181 Family Members Correlates with Reduced Expression of ATXN3 in Lymphocytes from SCA3 Patients[J].MicroRNA (Shariqah, United Arab Emirates), 2019, 8(1):76-85.

McLoughlin HS, Moore LR, Chopra R, et al. Oligonucleotide therapy mitigates disease in spinocerebellar ataxia type 3 mice. Ann Neurol. 2018 Jul;84(1):64-77.

Martier R , Sogorb-Gonzalez M , Stricker-Shaver J ,et al.Development of an AAV-Based MicroRNA Gene Therapy to Treat Machado-Joseph Disease - ScienceDirect[J].Molecular Therapy — Methods & Clinical Development, 2019.

系統作製戦略

The sequences from the ATG start codon to downstream of exon 11 of the mouse Atxn3 gene were replaced with the sequences from the ATG start codon to downstream of exon 11 of the human ATXN3 gene.

Figure 1. Gene editing strategy of B6-hATXN3 mice.

適用分野

Research on Machado-Joseph disease (MJD, SCA3);

Preclinical evaluation of ATXN3-targeted drugs.

検証 Data

1. Detection of human ATXN3 gene and mouse Atxn3 gene expression

RT-qPCR analysis reveals that the brain and lungs of B6-hATXN3 mice exclusively express the human ATXN3 gene, while the brain and lungs of wild-type mice exclusively express the mouse Atxn3 gene.

Figure 2. Expression of human ATXN3 gene and mouse Atxn3 gene in the brain and lungs of 6-week-old female wild-type mice (WT) and B6-hATXN3 mice. ND：Not detected.

2. Western Blot analysis of ATXN3 protein expression

Western blot analysis showed that the brains of wild-type mice expressed only murine ATXN3 protein (approximately 42 kDa), while the brains of B6-hATXN3 mice expressed only human ATXN3 protein (approximately 48 kDa)**.

Figure 3. Expression of ATXN3 protein in the brains of 6-week-old female wild-type mice and B6-hATXN3 mice. *The human ATXN3 gene and the murine Atxn3 gene are highly homologous. The human ATXN3 protein (361aa) has a slightly longer amino acid sequence than the murine ATXN3 protein (355aa). Although the predicted molecular weights of both the endogenous murine ATXN3 protein and the human ATXN3 protein are approximately 42 KD, the murine Atxn3 gene contains few or no CAG repeats, while the number of CAG repeats in the human ATXN3 gene in healthy individuals ranges from 12 to 44. Consequently, the longer polyQ structure in the human ATXN3 protein may slow down its electrophoretic migration speed in Western Blot detection, resulting in an observed band size larger than the predicted value ^[4].

3. Gait analysis (2-months-old)

（1）Stride Width (2-month-old)

Statistical comparisons were performed using unpaired t-tests; "ns" indicates no significant differences.

Figure 4. The forelimb and hindlimb stride width in hATXN3 models at 2 months of age during the gait analysis. Mouse numbers: B6-hATXN3 mice, 10 females and 10 males, total 20; Wild-type mice, 12 females and 12 males, total 24; the same applies below.

（2）Left Forelimb/Hindlimb Stride Length (2-month-old)

Statistical comparisons were performed using unpaired t-tests; "ns" indicates no significant differences.

Figure 5. The left forelimb and hindlimb stride length in hATXN3 models at 2 months of age during the gait analysis.

（3）Right Forelimb/Hindlimb Stride Length (2-month-old)

Statistical comparisons were performed using unpaired t-tests; "ns" indicates no significant differences.

Figure 6. The right forelimb and hindlimb stride length in hATXN3 models at 2 months of age during the gait analysis. Indications for stride width and stride length (Summary of Figures 4 to 6): At 2 months of age, hATXN3 displayed no significant differences in stride width or stride length comparable to WT mice, indicating stable balance in the gait pattern of hATXN3 models.

4. Rotarod Test: Latency to Fall (2-month-old)

Statistical comparisons were performed using Mann-Whitney tests; "ns" indicates no significant differences.

Figure 7. The latency to fall in hATXN3 models at 2 months of age during the rotarod test. Indications: No significant differences in latency to fall during the rotarod test were observed between hATXN3 models and WT mice at 2 months of age, suggesting preserved locomotor activity and coordination in hATXN3 models at 2 months of age.

5. Gait Test (4-month-old)

（1）Stride Width (4-month-old)

Statistical comparisons were performed using unpaired t-tests for normally distributed data and the Mann–Whitney test for non-normally distributed data; "ns" indicates no significant differences, ***p < 0.001.

Figure 8. The forelimb and hindlimb stride width in hATXN3 models at 4 months of age during the gait analysis.

（2）Left Forelimb/Hindlimb Stride Length (4-month-old)

Statistical comparisons were performed using unpaired t-tests for normally distributed data and the Mann–Whitney test for non-normally distributed data; "ns" indicates no significant differences.

Figure 9. The left forelimb and hindlimb stride length in hATXN3 models at 4 months of age during the gait analysis.

（3）Right Forelimb/Hindlimb Stride Length (4-month-old)

Statistical comparisons were performed using either an unpaired t-test or Welch’s t-test, depending on variance equality; "ns" indicates no significant differences.

Figure 10. The right forelimb/hindlimb stride length in hATXN3 models at 4 months of age during the gait analysis. Indications for stride width and stride length (Summary of Figures 8 to 10): At 4 months of age, hATXN3 displayed no significant differences in stride width or stride length comparable to WT mice, indicating stable balance in the gait pattern of hATXN3 models.

6. Rotarod Test: Latency to Fall (4-month-old)

Statistical comparisons were performed using unpaired t-tests for normally distributed data and the Mann–Whitney test for non-normally distributed data; "ns" indicates no significant differences.

Figure 11. The latency to fall in hATXN3 models at 4 months of age during the rotarod test. Indications: No significant variations in latency to fall during the rotarod test were detected between hATXN3 models and WT mice, suggesting unchanged locomotor activity and coordination in hATXN3 models at 4 months old.

7. Phenotypic Comparison of B6-hATXN3 and TG-hATXN3(85Q) Mice

a. B6-hATXN3 Mice (Catalog Number: C001398)

The B6-hATXN3 model at 2 and 4 months of age exhibited stable locomotor balance during gait analysis and the rotarod test, comparable to WT mice.

b. TG-hATXN3(85Q) Mice (Catalog Number: C001397)

Tg-hATXN3 (85Q) demonstrated progressive impairments in locomotor balance in both gait analysis and the rotarod test at 2 and 4 months of age.

B6-hATXN3 Mouse

基本情報

検証 Data

関連リソース

遺伝子名

遺伝子別名

NCBI ID

染色体

MGI ID

さらに

データシート

系統詳細

参考文献

系統作製戦略

適用分野

1. Detection of human ATXN3 gene and mouse Atxn3 gene expression

2. Western Blot analysis of ATXN3 protein expression

3. Gait analysis (2-months-old)

4. Rotarod Test: Latency to Fall (2-month-old)

5. Gait Test (4-month-old)

6. Rotarod Test: Latency to Fall (4-month-old)

7. Phenotypic Comparison of B6-hATXN3 and TG-hATXN3(85Q) Mice

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