Name: Tmem119-CreERT2 Mouse
Brand: サイヤジェン（Cyagen）
SKU: C001826

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Tmem119-CreERT2 Mouse

製品名

Tmem119-CreERT2 Mouse

製品ID

C001826

系統名

C57BL/6JCya-Tmem119^em2(CreERT2)/Cya

背景情報

C57BL/6JCya

Expressing Tissues/Cells

Microglia

状況

このマウス系統を論文で使用する場合は、「Tmem119-CreERT2 Mouse（カタログ番号C001826）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 TMEM119 gene encodes transmembrane protein 119 (TMEM119), also known as osteoblast-inducing factor (OBIF). It acts as an oncogene in osteosarcoma and is highly expressed in microglia. This ubiquitously expressed protein plays critical roles in various biological processes, including cell adhesion, migration, and differentiation. The TMEM119 gene encodes transmembrane protein 119 (TMEM119), also known as osteoblast-inducing factor (OBIF). It acts as an oncogene in osteosarcoma and is highly expressed in microglia. This ubiquitously expressed protein plays critical roles in various biological processes, including cell adhesion, migration, and differentiation. TMEM119 is vital for osteoblastogenesis—the differentiation of mesenchymal stem cells into bone-forming osteoblasts. It interacts with key signaling molecules like bone morphogenetic proteins (BMPs) to regulate osteoblast proliferation and differentiation, ultimately promoting bone formation and remodeling. Notably expressed in the developing nervous system, TMEM119 enhances neurite outgrowth and neuronal survival. It interacts with extracellular matrix components and growth factor signaling pathways to support the complex processes of neural network formation and function. Beyond its roles in bone formation and neurodevelopment, TMEM119 is also essential for normal spermatogenesis and late-stage testicular differentiation.

The Tmem119-CreERT2 mouse is generated via gene-editing technology by integrating a tamoxifen-inducible CreERT2 recombinase cassette into the stop codon of the murine Tmem119 gene, thereby driving its specific expression. This construction strategy ensures that the expression pattern of CreERT2 recombinase mimics that of the endogenous murine Tmem119 gene. Before tamoxifen induction, CreERT2 is confined to the cytoplasm; only after tamoxifen treatment can CreERT2 recombinase translocate into the nucleus to exert its genetic recombination function. When this strain is crossed with mice containing loxP sites, tamoxifen induction triggers Cre-mediated sequence recombination between loxP sites specifically in microglia. Tmem119-CreERT2 mice are applicable for researching the functions of resident microglia in the central nervous system, as well as bone development, osteoblast differentiation, and osteosarcoma.

系統作製戦略

The TAA stop codon was replaced with P2A-CreERT2 cassette.

Figure 1. Gene editing strategy of Tmem119-CreERT2 mice.

検証 Data

1. Method

Tmem119-CreERT2 mice were mated with Rosa26-LSL-tdTomato mice to generate double-allele heterozygous offspring. When the offspring reached a certain age (in weeks), they were induced via intraperitoneal injection of a specific dose of tamoxifen. CreERT2-mediated recombination, triggered by tamoxifen (Tam) induction, results in tdTomato protein expression in Cre-positive cells of the offspring. Seven days after completion of induction, brain tissues (hippocampus, cerebral cortex) of the offspring were collected, and Cre recombinase-mediated fluorescence was detected via immunofluorescence staining.

2. Group

Cre+Tam+ (G1): Tmem119-CreERT2[KI/+];Rosa26-LSL-tdTomato[CKI/+]; 5 day, 200 mg/kg/day (i.p)

Cre+Tam+ (G2): Tmem119-CreERT2[KI/+];Rosa26-LSL-tdTomato[CKI/+]; 5+5 day, 200 mg/kg/day (i.p)

Cre+Tam+ (G3): Tmem119-CreERT2[KI/+];Rosa26-LSL-tdTomato[CKI/+]; 5 day, 100 mg/kg/day (i.p)

Cre+Tam+ (G4): Tmem119-CreERT2[KI/+];Rosa26-LSL-tdTomato[CKI/+]; 5+5 day, 100 mg/kg/day (i.p)

Cre+Tam-: Tmem119-CreERT2[KI/+];Rosa26-LSL-tdTomato[CKI/+]; 5 day, Coin oil, (i.p)

Cre+Tam-: Tmem119-CreERT2[KI/+];Rosa26-LSL-tdTomato[CKI/+]; 5+5 day, Coin oil, (i.p)

Cre+: Tmem119-CreERT2[KI/+];Rosa26-LSL-tdTomato[CKI/+]; 5 day, 200 mg/kg/day (i.p)

*5+5 day: Intraperitoneal injection for 5 consecutive days, followed by a 1-day interval, then another 5 days of injection.

3. Result

（1）Expression of Cre recombinase in the hippocampus

① Histological analysis showed green fluorescent signals in the hippocampus of mice in the experimental groups (G1–G4), indicating Cre recombination had occurred in these tissues. A large number of red fluorescent signals were also detected in the hippocampus, with most overlapping with green fluorescence, confirming that Cre recombination occurred in IBA1+ microglia in the model.

② Satisfactory recombination efficiency was achieved under the G3 induction condition, while G2 showed slightly higher efficiency than the other three experimental groups. Therefore, the G2 induction protocol is recommended when experimental conditions permit.

③ Some green background fluorescence was observed in brain tissues, with non-specific staining detected in structures such as blood vessels. In the hippocampus of the control and blank groups, abundant red fluorescence and a small amount of green fluorescence were observed; the latter is presumed to be non-specific staining & background fluorescence.

Figure 2. Expression of Cre recombinase in the hippocampus after tamoxifen induction.

（2）Expression of Cre recombinase in the cerebral cortex

① Histological analysis revealed green fluorescent signals in the cerebral cortex of mice in the experimental groups (G1–G4), indicating Cre recombination had occurred in these tissues. A large number of red fluorescent signals were also detected in the cerebral cortex, with most overlapping with green fluorescence, confirming that Cre recombination occurred in IBA1+ microglia in the model.

③ Some green background fluorescence was observed in brain tissues, with non-specific staining detected in structures such as blood vessels. In the cerebral cortex of the control and blank groups, abundant red fluorescence and a small amount of green fluorescence were observed; the latter is presumed to be non-specific staining & background fluorescence.

Figure 3. Expression of Cre recombinase in the cerebral cortex after tamoxifen induction.

4. Summary

In the Tmem119-CreERT2 mouse model, the expression of Cre recombinase after tamoxifen induction is mainly localized to IBA1-expressing microglia, with good model specificity.

Recommended induction dose and sampling time: A continuous administration of tamoxifen (TAM) at a dose of 100 mg/kg/day for 5 days, followed by sampling 7 days after induction, can achieve satisfactory recombination efficiency. If experimental conditions permit, the induction dose can be appropriately increased to 200 mg/kg/day, or the induction duration can be adjusted to 10 days.

Abnormalities in tissues such as the ovary, spleen, and abdominal cavity may occur in Tmem119-CreERT2 mice after tamoxifen induction. Internal validation has not yet found a correlation between these abnormalities and variables such as induction duration, induction dose, sex, or genotype. The specific mechanism underlying these abnormalities remains unclear. Therefore, it is recommended to closely monitor abnormal manifestations in mice during the use of this model and adjust the experimental strategy on time.

Tmem119-CreERT2 Mouse

基本情報

検証 Data

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系統詳細

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4. Summary

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