Name: B6-htau Mouse
Brand: サイヤジェン（Cyagen）
SKU: C001410

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B6-htau Mouse

製品名

B6-htau Mouse

製品ID

C001410

系統名

C57BL/6JCya-Mapt^tm1(hMAPT)/Cya

背景情報

C57BL/6JCya

Note

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

状況

このマウス系統を論文で使用する場合は、「B6-htau Mouse（カタログ番号C001410）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

Small Nucleic Acids

製品タイプ

年齢

遺伝子型

性別

数量

お見積もりについてはこちらまでご連絡ください

HUGO-GT Humanized Models

Neurodegenerative Diseases

Small Nucleic Acids

基本情報

検証 Data

系統詳細

Frontotemporal Dementia (FTD) is the second most prevalent form of early-onset dementia, following Alzheimer’s disease (AD). This condition is distinguished by the selective degeneration of the frontal and temporal lobes, resulting in personality and behavioral changes, language impairments, and executive dysfunction. Approximately 40%-50% of FTD cases have a familial component, with known causative genes including MAPT, FUS, and TARDBP. Of these, MAPT is the earliest discovered and most frequently implicated in FTD. Mutations in the MAPT gene are detectable in roughly 30% of familial FTD cases^[1]. The tau protein, a microtubule-associated protein encoded by MAPT, is primarily localized to neuronal axons and plays a critical role in microtubule stability and assembly. By binding to microtubules, the tau protein helps to maintain neuronal cell shape. Mutations in MAPT can promote tau aggregation, leading to pathological tau protein accumulation and death of glutamatergic cortical neurons^[2]. Additionally, certain MAPT mutations can affect pre-mRNA exon splicing, altering the ratio of 3R to 4R tau protein isoforms and increasing the relative production of 4R-tau protein, which is more prone to fibril formation^[3-4].

Therapies targeting the MAPT gene primarily consist of small-molecule drugs and monoclonal antibodies, with indications including AD and FTD. Transgenic mice are frequently used in the drug development process, and the utilization of humanized animal models can facilitate the translation of promising treatments into clinical trials^[5-9].

This strain is a humanized mouse model in which the endogenous mouse Mapt gene has been replaced with its human counterpart, including the 3’UTR region. This model can be utilized to study various neurodegenerative diseases, such as FTD and AD. This model is commonly named htau. In addition, 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 the experimental needs in pharmacology and other fields.

参考文献

Bang J, Spina S, Miller BL. Frontotemporal dementia. Lancet. 2015 Oct 24;386(10004):1672-82.

Strang KH, Golde TE, Giasson BI. MAPT mutations, tauopathy, and mechanisms of neurodegeneration. Lab Invest. 2019 Jul;99(7):912-928.

Lisowiec J, Magner D, Kierzek E, Lenartowicz E, Kierzek R. Structural determinants for alternative splicing regulation of the MAPT pre-mRNA. RNA Biol. 2015;12(3):330-42.

Molecular Genetics Department, University of Antwerp. AD Mutations.

Andorfer C, Kress Y, Espinoza M, de Silva R, Tucker KL, Barde YA, Duff K, Davies P. Hyperphosphorylation and aggregation of tau in mice expressing normal human tau isoforms. J Neurochem. 2003 Aug;86(3):582-90.

Easton A, Jensen ML, Wang C, Hagedorn PH, Li Y, Weed M, Meredith JE, Guss V, Jones K, Gill M, Krause C, Brown JM, Hunihan L, Natale J, Fernandes A, Lu Y, Polino J, Bookbinder M, Cadelina G, Benitex Y, Sane R, Morrison J, Drexler D, Mercer SE, Bon C, Pandya NJ, Jagasia R, Ou Yang TH, Distler T, Grüninger F, Meldgaard M, Terrigno M, Macor JE, Albright CF, Loy J, Hoeg AM, Olson RE, Cacace AM. Identification and characterization of a MAPT-targeting locked nucleic acid antisense oligonucleotide therapeutic for tauopathies. Mol Ther Nucleic Acids. 2022 Aug 4;29:625-642.

DeVos SL, Miller RL, Schoch KM, Holmes BB, Kebodeaux CS, Wegener AJ, Chen G, Shen T, Tran H, Nichols B, Zanardi TA, Kordasiewicz HB, Swayze EE, Bennett CF, Diamond MI, Miller TM. Tau reduction prevents neuronal loss and reverses pathological tau deposition and seeding in mice with tauopathy. Sci Transl Med. 2017 Jan 25;9(374):eaag0481.

Yoshiyama Y, Higuchi M, Zhang B, Huang SM, Iwata N, Saido TC, Maeda J, Suhara T, Trojanowski JQ, Lee VM. Synapse loss and microglial activation precede tangles in a P301S tauopathy mouse model. Neuron. 2007 Feb 1;53(3):337-51.

Arvinas. (2021). Arvinas 2021 Investor Day Presentation.

Alnylam Pharmaceuticals, Inc. (2023). Microtubule associated protein tau (mapt) irna agent compositions and methods of use thereof (Patent No. WO2023049871). World Intellectual Property Organization.

Eli Lilly and Company. (2023). Mapt rna interference agents (Patent No. WO2023114700). World Intellectual Property Organization.

系統作製戦略

The sequences from the ATG start codon to downstream of the endogenous mouse Mapt gene were replaced with the sequences from the ATG start codon to downstream of the human MAPT gene. The humanized regions include 3'UTR.

Figure 1. Gene editing strategy for B6-htau mice.

適用分野

Research on Frontotemporal dementia (FTD);

Research on Alzheimer's disease (AD);

Research on other neurodegenerative diseases.

検証 Data

1. Detection of human MAPT gene expression

The RT-qPCR analysis results indicate significant expression of the human MAPT gene in both the brain and kidneys of B6-htau mice, with slightly higher expression in female mice compared to males. In contrast, WT mice do not express the human MAPT gene.

Figure 2. Human MAPT gene expression in the brain and kidney of 6-week-old wild-type mice (WT) and B6-htau mice. ND: Not detected

2. Detection of mouse Mapt gene expression

The RT-qPCR analysis results showed that the mouse Mapt gene was expressed in the brain and kidney of WT mice, while there was no expression of the mouse Mapt gene in B6-htau mice.

Figure 3. Mouse Mapt gene expression in the brain and kidney of 6-week-old wild-type mice (WT) and B6-htau mice.

3. Human Tau protein expression

A human Tau protein-specific antibody (HT7) was used for Western blot detection to evaluate the expression of human Tau protein in the mouse brain. The results showed that human Tau protein expression was successfully detected in the cerebral cortex and hippocampus of B6-htau mice, while no human Tau protein expression was observed in WT mice.

Figure 4. Western blot analysis of human Tau protein expression in the cerebral cortex and hippocampus of B6-htau mice and wild-type (WT) mice (7-week-old, female).

4. Behavioral Testing: Open Field Test

（1）3-month-old

Figure 5. The travel distance (A-C) and central area time ratio (D-F) for WT, B6-htau, B6-htau*P301L, and B6-htau*P301S mice in open field test. Data were analyzed using an ordinary one-way ANOVA; ns indicates no significant differences, **p < 0.01. (A-C) No significant alterations in travel distance across all models. (D-F) A notable decrease in the time spent in the central area is observed in B6-htau*P301S female when compared to their B6-htau counterparts, suggesting a slight increase in anxiety levels.

（2）6-month-old

Figure 6. The travel distance (A-C) and central area time ratio (D-F) for WT, B6-htau, B6-htau*P301L, and B6-htau*P301S mice in open field test. Data were analyzed using an ordinary one-way ANOVA; ns indicates no significant differences, *p < 0.05, **p < 0.01. (A-C) In comparison to WT mice, the B6-htau, B6-htau*P301S, and B6-htau*P301L exhibited a significant rise in travel distance. Given that no differences were noted among the three HUGO strains, we suggest that this increase in travel distance is likely a physiological characteristic rather than a pathological change (e.g., hyperactivity). (D-F) No significant alterations in the central area time ratio across all models.

（3）9-month-old

(A~C) No significant changes in travel distance were observed across all male models, indicating stable spontaneous locomotor activity.

(D~F) No significant variations in the time ratio spent in the central area were found across all models, suggesting unchanged anxiety levels.

Figure 7. The travel distance (A-C) and central area time ratio (D-F) for WT, B6-htau, B6-htau*P301L, and B6-htau*P301S mice in open field test. (A~C): The travel distance of 9-month-old mice over a duration of 10 minutes in an open field arena was measured. Statistical comparisons were performed using one-way ANOVA; ns indicates no significant differences. (D~F): The time ratio of the central area (central area time spent/total time spent) of the open field arena by 9-month-old mice was measured over a duration of 10 minutes. Statistical comparisons were performed using one-way ANOVA; ns indicates no significant differences.

5. Behavioral Testing: Rotarod Test

（1）3-month-old

No significant alterations in the latency across all models.

Figure 8. The latency for WT, B6-htau, B6-htau*P301L, and B6-htau*P301S mice in rotarod test. Data were analyzed using an ordinary one-way ANOVA; ns indicates no significant differences.

（2）6-month-old

No significant alterations in the latency across all models.

Figure 9. The latency for WT, B6-htau, B6-htau*P301L, and B6-htau*P301S mice in rotarod test. Data were analyzed using an ordinary one-way ANOVA; ns indicates no significant differences.

（3）9-month-old

No significant differences in latency for the rotarod assay were detected across all mixed-sex models, indicating consistent endurance and balance.

Figure 10. The latency observed in 9-month-old mice prior to their fall from the rotarod was measured. Statistical comparisons were conducted using one-way ANOVA for normally distributed data and the Kruskal–Wallis test for non-normally distributed data; ns indicates no significant differences.

6. Behavioral Testing: Novel Object Recognition Test

（1）6-month-old, Male

For the male comparison, WT, B6-htau, and B6-htau*P301L displayed a strong tendency to investigate new objects. In contrast, B6-htau*P301S mice exhibited almost no variation in their exploration patterns between familiar and novel objects, implying a considerable deficit in episodic memory.

Figure 11. The object preference (percentage of exploring the specific object/total exploration time of both objects) for male WT, B6-htau, B6-htau*P301L, and B6-htau*P301S mice in novel object recognition test. Data were analyzed using the t-test; ns indicates no significant differences, *p < 0.05, **p < 0.01.

（2）6-month-old, Female

For the female comparison, WT and B6-htau displayed a strong tendency to investigate new objects. In contrast, B6-htau*P301L mice showed a minor decrease in their preference for exploring new objects, resulting in an insignificant distinction between the familiar and novel subjects, indicating a slight impairment in episodic memory. Meanwhile, B6-htau*P301S mice exhibited almost no variation in their exploration patterns between familiar and novel objects, implying a considerable deficit in episodic memory.

Figure 12. The object preference (percentage of exploring the specific object/total exploration time of both objects) for female WT, B6-htau, B6-htau*P301L, and B6-htau*P301S mice in novel object recognition test. Data were analyzed using the t-test; ns indicates no significant differences, *p < 0.05, **p < 0.01.

（3）6-month-old, Male & Female

WT and B6-htau mice exhibited a strong tendency to investigate unfamiliar objects. In contrast, the previously observed lack of this inclination in B6-htau*P301L mice became evident in a mixed-sex setting, potentially due to the increased group size. Meanwhile, B6-htau*P301S mice showed minimal to no variation in their exploration patterns between familiar and novel objects, indicating a notable deficit in episodic memory.

Figure 13. The object preference (percentage of exploring the specific object/total exploration time of both objects) for mixed-sex WT, B6-htau, B6-htau*P301L, and B6-htau*P301S mice in novel object recognition test. Data were analyzed using the t-test; ns indicates no significant differences, *p < 0.05, ***p < 0.001.

（4）9-month-old, Male

Male WT and B6-htau mice demonstrated a strong preference for exploring the novel object, indicative of intact episodic memory. In contrast, male B6-htau*P301L and B6-htau*P301S mice showed little to no difference in exploration between familiar and novel objects, suggesting a marked impairment in episodic memory function.

Figure 14. The object preference (percentage of exploring the specific object/total exploration time of both objects) of 9-month-old male mice over a duration of 10 minutes was measured. Statistical comparisons were performed using a paired t-test; ns indicates no significant differences, *p < 0.05, ***p < 0.001.

（5）9-month-old, Female

Female WT and B6-htau mice exhibited a strong preference for exploring the novel object, reflecting intact episodic memory. In contrast, female B6-htau*P301S mice showed a slight, but statistically non-significant increase in preference for the novel object, suggesting a mild impairment in episodic memory. Female B6-htau*P301L mice displayed almost no difference in exploration between the familiar and novel objects, indicating a substantial deficit in episodic memory.

Figure 15. The object preference (percentage of exploring the specific object/total exploration time of both objects) of 9-month-old female mice over a duration of 10 minutes was measured. Statistical comparisons were performed using a paired t-test; ns indicates no significant differences, ***p < 0.001, ****p < 0.0001.

（6）9-month-old, Male & Female

Mixed-sex WT and B6-htau mice demonstrated a robust preference for exploring the novel object, indicative of preserved episodic memory. In contrast, B6-htau*P301L and B6-htau*P301S mice exhibited little to no difference in exploration between familiar and novel objects, suggesting a significant impairment in episodic memory.

Figure 16. The object preference (percentage of exploring the specific object/total exploration time of both objects) of 9-month-old mixed-sex mice over a duration of 10 minutes was measured. Statistical comparisons were performed using a paired t-test; ns indicates no significant differences, ***p < 0.001, ****p < 0.0001.

7. Validation of Pathological Changes for htau Strains

（1）9-month-old, B6-htau mice, HT7

HT7 is a monoclonal antibody that binds to an epitope present on all isoforms of human tau, enabling the visualization of overall tau expression. In contrast, AT8 selectively recognizes tau phosphorylated at Ser202 and Thr205, serving as a marker for pathological tau hyperphosphorylation commonly observed in Alzheimer's disease and related tauopathies.

Figure 17. Visualization of DAPI (blue), NeuN (red), and HT7 (green) staining in the hippocampus of C57BL/6J wild-type (WT) and B6-htau mice. Both C57BL/6J WT and B6-htau models displayed normal DAPI and NeuN staining, indicating proper nuclear morphology and well-organized neuronal architecture. No HT7 signal (green) was detected in the C57BL/6J WT mice, confirming the absence of human tau protein expression. In contrast, HT7 immunoreactivity (green) was evident in the B6-htau models, indicating the expression of human tau protein. HT7 staining was performed using a Thermo Fisher antibody (catalog number MN1000) at a dilution of 1:1,000.

（2）9-month-old, B6-htau*P301L and B6-htau*P301S mice, HT7

Figure 18. Visualization of DAPI (blue), NeuN (red), and HT7 (green) staining in the hippocampus of B6-htau*P301L and B6-htau*P301S models. In both models, DAPI staining (blue) reveals abnormal neuronal nuclear morphology, while NeuN immunoreactivity (red) indicates disorganized neuronal architecture. HT7 immunoreactivity (green) confirms the expression of human tau protein in the hippocampal tissue. HT7 staining was conducted using a Thermo Fisher antibody (catalog number MN1000) at a dilution of 1:1,000.

（3）9-10-month-old, B6-htau mice, AT8

Figure 19. Visualization of DAPI (blue), NeuN (red), and AT8 (yellow) staining in the hippocampus of C57BL/6J WT and B6-htau models. Normal DAPI and NeuN staining were observed in both models, indicating intact nuclear morphology and organized neuronal arrangement. The absence of the AT8 signal suggests no phosphorylated tau protein, indicating the lack of pathological tau in the C57BL/6J WT and B6-htau mice. AT8 staining was performed using a Thermo Fisher antibody (catalog number MN1020) at a dilution of 1:1,000.

（4）9-month-old, B6-htau*P301L and B6-htau*P301S mice, AT8

Figure 20. Visualization of DAPI (blue), NeuN (red), and AT8 (yellow) staining in the hippocampus of B6-htau*P301L and B6-htau*P301S models. In both B6-htau*P301L and B6-htau*P301S models, DAPI staining (blue) depicts abnormal neuronal nuclear morphology, while NeuN immunoreactivity (red) illustrates a disorganized neuronal architecture. AT8 immunoreactivity (yellow) confirms the presence of phosphorylated human tau protein, indicating pathological tau in these models. AT8 staining was performed using a Thermo Fisher antibody (catalog number MN1020) at a dilution of 1:1,000.

8. Small interfering RNA (siRNA) drugs can significantly reduce the expression of human MAPT mRNA in B6-htau mice.

（1）Case I

Figure 21. Expression of human MAPT gene in the hippocampus and frontal cortex of 14-week-old male homozygous B6-htau mice after treatment with siRNA drug candidate molecules (AD-1637701 and Conjugate 31*)**. Following a single intracerebroventricular (icv) injection of AD-1637701 and Conjugate 31, hippocampus and frontal cortex samples were collected on day 14 for RT-qPCR analysis. The result shows a significant reduction in human MAPT mRNA expression in the hippocampus and frontal cortex of B6-htau mice treated with AD-1637701 and Conjugate 31 (data presented as mean±SD). *AD-1637701 and Conjugate 31 are siRNA drug candidate molecules from disclosed patents^[10-11]. **Data provided by the client.

（2）Case II

Figure 22. Significant reduction in human MAPT gene expression in the thoracic spinal cord, cerebellum, hippocampus, and lumbar spinal cord of 8-week-old female homozygous B6-htau mice following treatment with candidate siRNAs targeting human MAPT (siRNA1, siRNA2, siRNA3)*. After a single intracerebroventricular (icv) injection of siRNA molecules, the thoracic spinal cord, cerebellum, hippocampus, and lumbar spinal cord of the mice were collected on day 28 for RT-qPCR analysis. The results show that after siRNA drug treatment, the expression of human MAPT mRNA was significantly reduced in the thoracic and lumbar spinal cord, hippocampus, and cerebellum of B6-htau mice. (Bars represent mean ± SEM, n=3) *Data provided by the client.

B6-htau 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 MAPT gene expression

2. Detection of mouse Mapt gene expression

3. Human Tau protein expression

4. Behavioral Testing: Open Field Test

5. Behavioral Testing: Rotarod Test

6. Behavioral Testing: Novel Object Recognition Test

7. Validation of Pathological Changes for htau Strains

8. Small interfering RNA (siRNA) drugs can significantly reduce the expression of human MAPT mRNA in B6-htau mice.

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