Name: Cyp19a1-IRES-Cre Mouse
Brand: サイヤジェン（Cyagen）
SKU: C001382

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Cyp19a1-IRES-Cre Mouse

製品名

Cyp19a1-IRES-Cre Mouse

製品ID

C001382

系統名

C57BL/6JCya-Cyp19a1^{tm1(IRES-Cre)}/Cya

背景情報

C57BL/6JCya

状況

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Cre Mouse Models

基本情報

検証 Data

系統詳細

The CYP19A1 gene encodes an aromatase that is a member of the cytochrome P450 superfamily, also known as estrogen synthase, an enzyme responsible for a key step in estrogen biosynthesis. Cytochrome P450 proteins are monooxygenases that catalyze many reactions involved in drug metabolism and the synthesis of cholesterol, steroids, and other lipids. This protein is located in the endoplasmic reticulum and catalyzes the final step in estrogen biosynthesis. The CYP19A1 aromatase is responsible for aromatizing androgens into estrogens and can be found in many tissues, including the gonads (granulosa cells), brain, adipose tissue, placenta, blood vessels, skin, and bone, as well as tissues from endometriosis, uterine fibroids, breast cancer, and endometrial carcinoma. It is an important factor in sexual development ^[1-2].

The Cyp19a1-IRES-Cre mice were constructed using gene editing technology to insert an IRES-Cre element downstream of the stop codon of the endogenous Cyp19a1 gene in mice. The expression pattern of Cre recombinase is similar to that of the endogenous gene. When this strain is crossed with mice containing loxP sites, sequence recombination between loxP sites mediated by Cre recombinase can occur in the granulosa cells of the ovaries of offspring mice.

参考文献

Manikandan P, Nagini S. Cytochrome P450 Structure, Function and Clinical Significance: A Review. Curr Drug Targets. 2018;19(1):38-54.

Azcoitia I, Mendez P, Garcia-Segura LM. Aromatase in the Human Brain. Androg Clin Res Ther. 2021 Dec 23;2(1):189-202.

系統作製戦略

The “IRES-Cre” cassette was inserted downstream of the TGA stop codon.

Figure 1. Diagram of the gene editing strategy for the generation of Cyp19a1-IRES-Cre mice.

検証 Data

1. Method

The Cyp19a1-IRES-Cre mice were bred with Rosa26-LSL-tdTomato mice to generate double heterozygous offspring. Cre-mediated recombination will result in the expression of tdTomato protein in the Cre-positive cells of the offspring. The ovaries, testes, and brain tissue of 6-week-old offspring mice were collected and immunofluorescence was used to observe the expression of tdTomato protein to determine the activity of Cre recombinase.

2. Genotype

Cre+: Cyp19a1-IRES-Cre[KI/+]; Rosa26-LSL-tdTomato[CKI/+]

Cre-: Rosa26-LSL-tdTomato[CKI/+]

3. Result

（1）Expression of Cre recombinase in the female gonads

In Cre+ female mice, there was a large amount of red fluorescence from the tdTomato protein in the ovarian granulosa cells, indicating a large amount of Cre recombinase expression in this location. In contrast, no expression of tdTomato red fluorescence was detected in the control group (Cre-) mice, indicating the absence of Cre recombinase.

Figure 2. Immunofluorescence staining of the ovaries in female mice.

（2）Expression of Cre recombinase in the male gonads

In Cre+ male mice, there was red fluorescence in a small number of testicular cells, which partially overlapped with DAPI staining, indicating that these cells may be testicular support cells or other types of cells. In contrast, no expression of tdTomato red fluorescence was detected in the control group (Cre-) mice, indicating the absence of Cre recombinase.

Figure 3. Immunofluorescence staining of the testis in male mice.

（3）Expression of Cre recombinase in the hippocampal region of the brain

In Cre+ mice, there was partial red fluorescence in the hippocampal CA region* (as shown within the white box in the image), indicating that Cre recombination has occurred in this area. In contrast, no expression of tdTomato red fluorescence was detected in the control group (Cre-) mice, indicating the absence of Cre recombinase.

Figure 4. Immunofluorescence staining of the hippocampal region of the brain. *The hippocampal CA region refers to the Cornu Ammonis region of the hippocampal formation, which includes areas CA1 and CA2.

（4）Expression of Cre recombinase in the ventricles, fiber tracts, and nucleus accumbens regions of the brain

In Cre+ mice, there was partial red fluorescence in the lateral ventricles*, stria terminalis*, and caudoputamen structures*, indicating that Cre recombination has occurred in these areas. In contrast, no expression of tdTomato red fluorescence was detected in the control group (Cre-) mice, indicating the absence of Cre recombinase.

Figure 5. Immunofluorescence staining of the ventricles, fiber tracts, and nuclei of the brain. *The fiber tract region: stria terminalis (as shown by the left arrow in the image). *The ventricular region: lateral ventricle (as shown by the middle arrow in the image). *The brain nuclei region: caudoputamen (as shown by the right arrow in the image).

4. Summary

In the Cyp19a1-IRES-Cre mice, the expression of Cre recombinase is mainly localized in the ovarian granulosa cells. At the same time, recombination mediated by Cre recombinase can also occur in some cells in the hippocampus, ventricles, fiber tracts, and brain nuclei of the brain. The expression of Cre recombinase is similar to the expression pattern of the endogenous Cyp19a1 gene in mice and has good specificity.

Cyp19a1-IRES-Cre Mouse

基本情報

検証 Data

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系統詳細

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系統作製戦略

1. Method

2. Genotype

3. Result

4. Summary

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