Name: huC3 Mouse
Brand: サイヤジェン（Cyagen）
SKU: C001955

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huC3 Mouse

製品名

huC3 Mouse

製品ID

C001955

系統名

C57BL/6JCya-C3^tm1(hC3)/Cya

背景情報

C57BL/6JCya

状況

このマウス系統を論文で使用する場合は、「huC3 Mouse（カタログ番号C001955）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

Cytokine Gene Humanized Mouse Models

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遺伝子型

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HUGO-GT Humanized Models

Cytokine Gene Humanized Mouse Models

基本情報

検証 Data

系統詳細

Complement component C3 plays a central role in activating the complement system and is the most abundant complement protein in human plasma, primarily synthesized in the liver. As part of the innate immune system, the complement system is activated during tissue damage and pathogen invasion, playing a crucial role in the inflammatory response, host homeostasis, and pathogen defense. The complement cascade is activated through the classical pathway, alternative pathway, and lectin pathway, all of which generate C3 convertase, which cleaves C3 into C3a and C3b. C3a is a potent anaphylatoxin with pro-inflammatory activity, while C3b is a regulator that induces C5 cleavage, thereby participating in the dissolution and clearance of immune complexes. Mutations in this gene are associated with atypical hemolytic uremic syndrome (aHUS) and age-related macular degeneration (AMD). Deficiencies in C3 and C3-derived peptides can lead to autoimmune diseases (such as rheumatoid arthritis, systemic lupus erythematosus, and vasculitis) and make individuals susceptible to recurrent respiratory infections and infections caused by encapsulated organisms. Conversely, excessive activation of C3 and related complement components is associated with kidney diseases (immune complex glomerulonephritis, hemolytic uremic syndrome, lupus nephritis, membranous nephropathy, and immune-mediated nephropathy) ^[1-2].

The huC3 mouse is a mouse C3 humanized model created by replacing the mouse C3 gene with the human C3 gene using gene-editing technology. The humanized regions include the 5’UTR and 3’UTR. Under natural breeding conditions, homozygous huC3 mice exhibit mortality around 10 weeks of age, and it has been observed that heterozygous mice also experience mortality. Additionally, based on the innovative TurboKnockout technology combined with BAC recombination developed by Cyagen Biosciences, customized services are available for different mutations to meet the experimental needs of researchers studying complement-mediated diseases.

参考文献

Delanghe JR, Speeckaert R, Speeckaert MM. Complement C3 and its polymorphism: biological and clinical consequences. Pathology. 2014 Jan;46(1):1-10.

Yates JR, Sepp T, Matharu BK, Khan JC, Thurlby DA, Shahid H, Clayton DG, Hayward C, Morgan J, Wright AF, Armbrecht AM, Dhillon B, Deary IJ, Redmond E, Bird AC, Moore AT; Genetic Factors in AMD Study Group. Complement C3 variant and the risk of age-related macular degeneration. N Engl J Med. 2007 Aug 9;357(6):553-61.

系統作製戦略

The sequences from upstream of exon 1 to the TGA stop codon of mouse C3 were replaced with the sequences from upstream of exon 1 to downstream of exon 41 of human C3.

Figure 1. Gene editing strategy of huC3 mice.

適用分野

Preclinical research on C3-targeted drugs;

Research in immunotherapy, oncology, etc.

検証 Data

1. RT-qPCR

RT-qPCR results showed that human C3 mRNA was expressed in the eyes, kidneys, livers, lungs, and spleens of 5-week-old huC3 mice, while mouse C3 mRNA was not expressed (n=4, data are presented as mean±SD).

Figure 2. C3 gene expression in various tissues of wild-type (WT) and huC3 mice.

2. ELISA

ELISA results showed that human C3 protein was abundantly detected in the serum of homozygous male, homozygous female, and heterozygous female huC3 mice. (huC3 homozygous and WT mice: 5 weeks old; huC3 heterozygous mice: 7 weeks old; n=5; data presented as mean ± SD.)

Figure 3. Expression of human C3 protein in the serum of wild-type (WT) and huC3 mice.

3. Western Blot

Western Blot results showed successful expression of human C3 protein in the livers of huC3 mice, while no human C3 protein was expressed in the livers of wild-type mice.

Figure 4. Expression of human C3 protein in the liver of wild-type (WT) and huC3 mice.

4. IHC Staining (10 & 16 weeks)

IHC validation showed that human C3 expression was present in the liver of huC3 mice at both 10 and 16 weeks of age (red arrows indicate positive signals), while no positive signals were detected in WT mice of the corresponding ages.

Figure 13a. Liver immunohistochemistry (IHC) results of 10-week-old male wild-type (WT) mice and homozygous huC3 mice.

Figure 13b. Liver immunohistochemistry (IHC) results of 16-week-old male wild-type (WT) mice and homozygous huC3 mice.

5. Retinal phenotypes in WT and huC3 mice

The fundus morphology and retinal OCT results of homozygous huC3 mice were consistent with those of wild-type mice.

Figure 5. Fundus morphology and OCT results of WT and huC3 mice.
*OD: Oculus Dexter; OS: Oculus Sinister

6. Electroretinogram (ERG)

Compared with WT, the amplitudes of the a-wave and b-wave in both scotopic and photopic ERG recordings of homozygous huC3 mice were nearly identical to those of the WT. The retinal photoreceptor function of homozygous huC3 mice were normal.

Figure 6. Electroretinogram (ERG) detection results of WT and huC3 mice at 8 weeks old.

7. Kidney Histopathology

（1）H&E Staining (10 weeks)

Figure 7. H&E staining results of kidneys from 10-week-old male wild-type (WT) and homozygous huC3 mice. Legend: Glomerulosclerosis (yellow arrow), columnar epithelial cells of the Bowman's capsule parietal layer (green arrow), tubular atrophy (orange arrow), tubular dilation (yellow polygon), protein casts (green polygon), fibrous connective tissue proliferation (cyan arrow), granulocyte (blue arrow) and lymphocyte (silver arrow) infiltration.

（2）H&E Staining (16 weeks)

Figure 8. H&E staining results of kidneys from 16-week-old male wild-type (WT) and homozygous huC3 mice.
Legend: Glomerulosclerosis (yellow arrow), columnar epithelial cells of the Bowman's capsule parietal layer (green arrow), tubular atrophy (orange arrow), tubular dilation (yellow polygon), fibrous connective tissue proliferation (cyan arrow), minimal granulocyte (blue arrow) and lymphocyte (silver arrow) infiltration, and necrotic cell debris (brown arrow).

（3）PAS Staining (10 weeks)

Figure 9. PAS staining results of kidneys from 10-week-old male wild-type (WT) and homozygous huC3 mice.
Legend: Glomerular mesangial matrix proliferation (yellow arrow) and thickening of the Bowman's capsule basement membrane (green arrow).

（4）PAS Staining (16 weeks)

Figure 10. PAS staining results of kidneys from 16-week-old male wild-type (WT) and homozygous huC3 mice.
Legend: Glomerular mesangial matrix proliferation (yellow arrow) and thickening of the Bowman's capsule basement membrane (green arrow).

（5）Summary and Analysis of Kidney Histopathology

Statistical data showed that the glomeruli, tubules, and interstitium of WT mice appeared normal. In contrast, the kidneys of huC3 mice exhibited various pathological changes, including glomerulosclerosis, columnar epithelial cells of the Bowman's capsule parietal layer, tubular atrophy and dilation, and protein cast formation. Furthermore, the renal interstitium showed fibrous connective tissue proliferation with granulocyte and lymphocyte infiltration. The pathological scores for glomeruli, tubules, and interstitium in huC3 mice were significantly higher than those in WT mice.

Figure 11. Pathological analysis of kidneys from wild-type (WT) and homozygous huC3 mice.

8. Liver Histopathology and Analysis

（1）H&E Staining (10 & 16 weeks)

Most wild-type mice showed regularly and neatly arranged hepatic cords with no significant expansion or compression of the sinusoids. Occasional hepatocyte vacuolar degeneration, focal granulocyte infiltration within the hepatic lobule, and occasional focal lymphocyte infiltration around the portal area were observed, with no evident fibrosis. In contrast, huC3 mice showed large amounts of mild to moderate hepatocyte edema, with swollen cells and loose, pale cytoplasm. The hepatic sinusoids were compressed, and the hepatic cords were poorly arranged. Occasional focal granulocyte infiltration was seen around the portal area, and a small amount of fibrous connective tissue proliferation was observed around the portal area and in the hepatic sinusoids in some mice.

Figure 12a. H&E staining results of kidneys from 10-week-old male wild-type (WT) and homozygous huC3 mice.
Legend: Mild hepatocyte edema (yellow arrow), hepatocyte vacuolar degeneration (cyan arrow), focal granulocyte infiltration (green arrow), occasional focal lymphocyte infiltration around the portal area (blue arrow).

Figure 12b. H&E staining results of kidneys from 16-week-old male wild-type (WT) and homozygous huC3 mice.
Legend: Mild hepatocyte edema (yellow arrow), hepatocyte vacuolar degeneration (cyan arrow), focal granulocyte infiltration (green arrow), occasional focal lymphocyte infiltration around the portal area (blue arrow).

9. Growth and Survival Curves

Growth curve data showed that the body weight of homozygous huC3 male mice was lower than that of wild-type mice, while there was no significant difference in females (statistical analysis method: Two-ANOVA). Survival curve data showed that the first death in homozygous huC3 male mice occurred at day 85, with 100% mortality by day 196. The first death in homozygous huC3 female mice occurred at day 78, with a 60% survival rate by day 274 (statistical analysis method: Log-rank (Mantel-Cox) test).

Figure 14. Comparison of growth and survival curves for wild-type (WT) and homozygous huC3 mice.

huC3 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. RT-qPCR

2. ELISA

3. Western Blot

4. IHC Staining (10 & 16 weeks)

5. Retinal phenotypes in WT and huC3 mice

6. Electroretinogram (ERG)

7. Kidney Histopathology

8. Liver Histopathology and Analysis

9. Growth and Survival Curves

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