Name: huC3 Mouse
Brand: 싸이아젠(Cyagen)
SKU: C001955

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huC3 Mouse

제품명

huC3 Mouse

제품 ID

C001955

품종 계통

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

Backgroud

C57BL/6JCya

상태

이 마우스 계통을 논문에서 사용할 경우, “huC3 Mouse (카탈로그 번호 C001955)은 Cyagen에서 구입하였습니다.”라고 명시해 주시기 바랍니다.

HUGO-GT Humanized Models

Cytokine Gene Humanized Mouse Models

구매 가능한 제품 종류

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Genotype

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가격 문의

HUGO-GT Humanized Models

Cytokine Gene Humanized Mouse Models

기본 정보

검증 데이터

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.

Reference

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.

변형 전략

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.

응용 분야

Preclinical research on C3-targeted drugs;

Research in immunotherapy, oncology, etc.

검증 데이터

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

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Datasheet

품종 계통 설명

Reference

변형 전략

응용 분야

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

관련 제품

출판물