Name: Vil1-MerCreMer Mouse
Brand: サイヤジェン（Cyagen）
SKU: C001433

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Vil1-MerCreMer Mouse

製品名

Vil1-MerCreMer Mouse

製品ID

C001433

系統名

C57BL/6JCya-Igs2^{em1(Vil1-MerCreMer)}/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、「Vil1-MerCreMer Mouse（カタログ番号C001433）はサイアジェンから購入しました。」と引用してください。

Inducible Cre Mouse Models

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Inducible Cre Mouse Models

基本情報

検証 Data

系統詳細

The VIL1 gene encodes the Villin 1 protein, which belongs to the calcium-regulated actin-binding protein family and is a major part of the brush border cell cytoskeleton, playing a role in actin filament capping, severing, and bundling. Villin 1 is an epithelial cell-specific Ca(2+)-regulated actin-modifying protein that regulates the reorganization of microvillar actin filaments, playing a role in actin nucleation, actin filament bundle assembly, actin filament capping, and severing. VIL1 is selectively expressed in epithelial cells in the gastrointestinal tract, gallbladder, and renal tubules.

Vil1-MerCreMer mice were generated by inserting a Tamoxifen-inducible Cre recombinase protein MerCreMer gene expression element controlled by the mouse Vil1 promoter into the mouse H11 safe harbor site. Before induction, MerCreMer is only present in the cytoplasm and can only enter the nucleus to exert its recombination effect after Tamoxifen treatment. When bred with mice containing a loxP site-flanked sequence, Cre recombinase-mediated deletion of the flanked sequence will occur in the villous and crypt epithelial cells of the small and large intestines of the offspring after Tamoxifen induction. The onset of endogenous mouse Vil1 gene expression is at 9.0dpc, and the onset of Cre recombinase expression in this strain is approximately 12.5dpc.

系統作製戦略

The “Mouse Vil1 promoter-MerCreMer-rBG pA” gene expression element was inserted into the H11 locus.

Figure 1. Schematic diagram of gene editing in Vil1-MerCreMer mice.

検証 Data

1. Method

Vil1-MerCreMer mice were crossed with Rosa26-LSL-tdTomato mice to generate double heterozygous offspring. At 6 weeks of age, these offspring were subjected to Tamoxifen induction via intraperitoneal injection at a dose of 75 mg/kg for 5 consecutive days. This resulted in Cre recombinase-mediated deletion of LSL elements and subsequent expression of tdTomato in Cre-positive cells. One week after induction, tissues from the intestines, stomach, ureters, kidneys, ovaries, uterus, epididymis, and testes were collected from the offspring. The distribution of tdTomato was determined by immunofluorescence staining to assess the expression of Cre recombinase.

2. Groups

Cre+Tam+: Vil1-MerCreMer[KI/+];Rosa26-LSL-tdTomato[CKI/+], with Tamoxifen;

Cre+Tam-: Vil1-MerCreMer[KI/+];Rosa26-LSL-tdTomato[CKI/+], without Tamoxifen.

3. Result

（1）Expression of Cre recombinase in the small intestinal tissues

The presence of abundant tdTomato-positive signals in the crypts and intestinal villus epithelial cells of the duodenum, jejunum, and ileum in Cre+Tam+ mice indicates strong expression of Cre recombinase in the small intestine. In contrast, no tdTomato expression or Cre recombinase activity was observed in these tissues in the non-tamoxifen-induced group.

Figure 2. Immunofluorescence staining of duodenal, jejunal, and ileal tissues.

（2）Expression of Cre recombinase in the large intestinal tissues

The presence of abundant tdTomato-positive signals in the crypts and intestinal villus epithelial cells of the cecum, colon, and rectum in Cre+Tam+ mice indicates strong expression of Cre recombinase in the large intestine.

Figure 3. Immunofluorescence staining of the cecum, colon, and rectum tissues.

（3）Expression of Cre recombinase in the stomach, ureter, and kidney

The presence of a small amount of tdTomato-positive signal in kidney and stomach cells of Cre+Tam+ mice indicates trace expression of Cre recombinase. However, no such expression was observed in the ureter.

Figure 4. Immunofluorescence staining of the gastric, ureteric, and renal tissues.

（4）Expression of Cre recombinase in the female gonads

The absence of tdTomato signals in the uterus and ovaries of Cre+Tam+ mice suggests that there is no Cre recombinase activity at these sites, and therefore no germline deletion of the target gene.

Figure 5. Immunofluorescence staining of the uterus and ovary.

（5）Expression of Cre recombinase in the male gonads

The presence of only minimal amounts of tdTomato-positive signal in the testis and epididymis of Cre+Tam+ mice suggests that the likelihood of Cre recombinase expression leakage at these sites is extremely low, and therefore does not result in germline deletion of the target gene.

Figure 6. Immunofluorescence staining of the testicular and epididymal tissues.

4. Summary

In Vil1-MerCreMer mice, the expression of Cre recombinase was primarily localized to the crypts and intestinal villus epithelial cells of both the large and small intestines. Only a small amount of Cre recombinase activity was detected in certain cells of the kidney and stomach, while no expression was observed in the ureters. Furthermore, Cre recombinase activity was virtually absent in the gonads, indicating a low likelihood of germline leakage. This model represents a highly tamoxifen-inducible, tissue-specific Cre mouse line targeting intestinal crypts and intestinal villus epithelial cells.

Vil1-MerCreMer Mouse

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