The P2A-3xSV40 NLS-mScarlet cassette was inserted upstream of TGA stop codon. Nuclear-localized mScarlet is expressed under the regulatory control of Alpl gene elements in this mouse model. This localization is achieved through the Nuclear Localization Signal (NLS), which efficiently targets mScarlet to the cell nucleus. This enables mScarlet fluorescence protein nuclear tracing studies.

The ADIPOQ gene-encoded adiponectin is a protein hormone produced exclusively by adipocytes (fat cells). It is transported through the bloodstream to muscle and liver cells. Adiponectin regulates various pathways related to fat storage and metabolism, including the modulation of blood glucose levels, fatty acid breakdown, brown adipocyte differentiation, and negative regulation of gluconeogenesis. By increasing insulin sensitivity and promoting fatty acid breakdown, adiponectin plays a crucial role in regulating glucose and fat metabolism. Additionally, it exhibits direct anti-diabetic, anti-atherosclerotic, and anti-inflammatory activities ^[1-2]. The mutation of the ADIPOQ gene is associated with adiponectin deficiency syndrome. Although the ADIPOQ gene is expressed predominantly (or almost exclusively) in adipose tissue, adiponectin, as a secreted hormone, circulates via the bloodstream and is widely distributed in various tissues and organs, including skeletal muscle, liver, intestine, male reproductive glands, and brain, where it exerts its physiological effects through specific receptors (such as AdipoR1 and AdipoR2) ^[3-4]. The Adipoq-P2A-iCre mice are constructed by inserting a codon-improved Cre recombinase (iCre) element into the endogenous Adipoq gene of mice. The expression pattern of iCre recombinase is similar to the endogenous gene. When this strain is crossed with mice containing loxP sites, sequence recombination mediated by the Cre recombinase between loxP sites can occur in the white adipose tissue (WAT) and brown adipose tissue (BAT) of its offspring.

The TAA stop codon was replaced with the P2A-EGFP-T2A-iCre cassette. Cre recombinase and enhanced green fluorescent protein (EGFP) are expressed under the regulatory control of Alas2 gene elements.

Adiponectin, a protein hormone encoded by the ADIPOQ gene, is exclusively produced by adipocytes. It is secreted into the bloodstream and transported to muscle and liver cells, playing a crucial role in maintaining glucose and lipid metabolic homeostasis. Adiponectin participates in the regulation of glucose metabolism, lipolysis, and energy balance by promoting fatty acid oxidation, enhancing insulin sensitivity, and modulating metabolic signaling pathways such as AMPK. Additionally, it exhibits biological functions including anti-inflammatory and anti-atherosclerotic effects, as well as the amelioration of insulin resistance ^[1-3]. Aberrant ADIPOQ expression is closely associated with various metabolic disorders, including obesity, type 2 diabetes, metabolic syndrome, fatty liver disease, and cardiovascular diseases ^[1-4]. Although the ADIPOQ gene is expressed exclusively in adipose tissue, adiponectin is widely distributed across multiple organs, including muscle, liver, intestine, male reproductive glands, and the brain ^[3-4]. The Adipoq-P2A-iCreERT2 mouse was generated by inserting a P2A-iCreERT2 expression cassette at the stop codon of the endogenous mouse Adipoq gene. The regulatory elements of the mouse Adipoq gene drive the expression of the iCreERT2 recombinase. In the absence of tamoxifen, the iCreERT2 recombinase is predominantly retained in the cytoplasm; upon tamoxifen induction, the recombinase translocates into the nucleus to exert its recombinase activity. When Adipoq-P2A-iCreERT2 mice are crossed with mice carrying loxP sites, the resulting offspring are expected to undergo Cre-mediated recombination of sequences flanked by loxP sites within adipose tissue following tamoxifen induction.

For the Kl model, the TGA stop codon will be replaced by P2A-Cre. A synonymous mutation p.R894=(CGG to CGC) and an additional mutation c.*3C>G in 3'UTR will also be introduced to prevent the binding and re-cutting of the sequence.

The Cre-P2A cassette was inserted upstream of the ATG start codon.

The TAA stop codon was replaced with the P2A-CreERT2 cassette. CreERT2 recombinase is expressed under the regulatory control of Adgre1 gene elements. This model is a Tamoxifen-inducible Cre mouse, and when crossed with mice containing loxP sites, the offspring mice are expected to undergo sequence recombination between loxP sites mediated by Cre recombinase in macrophages following Tamoxifen induction.

The ADGRL2 gene (Adhesion G Protein-Coupled Receptor L2), also known as latrophilin 2, encodes a member of the adhesion G protein-coupled receptor (aGPCR) family, which are characterized by a long N-terminal domain involved in cell-cell and cell-matrix interactions ^[1]. The encoded protein, ADGRL2, is involved in various physiological processes, including cell adhesion, neuronal development, regulation of exocytosis (e.g., as a low-affinity receptor for alpha-latrotoxin), and maintaining intestinal homeostasis ^[2]. It is expressed in numerous tissues, with notable expression in the central nervous system (neurons, hippocampus), intestinal epithelium, and specifically, its expression is strongly upregulated during keratinocyte differentiation in epidermal tissue ^[3]. Dysregulation or variations in ADGRL2 have been associated with a range of conditions, including neurodegenerative diseases (like Alzheimer's and Parkinson's), inflammatory bowel diseases (Crohn's disease, ulcerative colitis), certain autoimmune diseases (rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis), and even metabolic syndrome and cocaine use disorder. Adgrl2-3xGGGGS-mCherry mice are constructed by replacing the partial exon 1 coding region of the mouse Adgrl2 gene with HA signal peptide - HA tag - Mouse Adgrl2 CDS (without signal peptide) - 3xGGGGS - mCherry - rBG pA cassette using gene editing technology. The Adgrl2-3xGGGGS-mCherry mouse carries a red fluorescent protein (mCherry) expression cassette, making it a precise research model that maintains protein function while offering fluorescence visualization. This model is valuable for several key areas of study. For instance, it can be used for the spatio-temporal dynamic analysis of the Adgrl2 gene expression profile. Researchers can also utilize it for investigating neuronal development and synapse formation mechanisms. Furthermore, it enables live-animal dynamic tracking and real-time imaging observation, providing invaluable insights. Lastly, this model is well-suited for systematic studies of protein interaction networks and downstream signaling pathways.

The AGRP gene encodes Agouti-related protein (AgRP), a neuropeptide synthesized by AgRP/NPY neurons predominantly located in the arcuate nucleus of the hypothalamus, as well as in the kidneys and adrenal glands. The expression of AGRP is modulated by various factors, including nutritional status and hormonal signals. Notably, AGRP expression is markedly upregulated during periods of starvation and rapidly downregulated following refeeding. AgRP is exclusively synthesized in the ventromedial part of the arcuate nucleus within neuropeptide Y (NPY)-containing cells, where it is co-expressed with NPY. This neuropeptide plays a pivotal role in enhancing appetite, reducing metabolic rate, and decreasing energy expenditure, making it one of the most potent and enduring appetite stimulators. AgRP exerts its orexigenic effects by antagonizing melanocortin receptor 4 (MC4R), thereby promoting food intake and inhibiting energy expenditure, which is crucial for weight regulation. Mutations in the AGRP gene have been implicated in conditions such as late-onset obesity and anorexia nervosa, underscoring its significant role in energy homeostasis and body weight control. The Agrp-IRES-CreERT2-P2A-tdTomato mouse model was generated by integrating the IRES-CreERT2-P2A-tdTomato gene expression cassette into the endogenous Agrp locus via gene editing technology. Under the control of the mouse endogenous Agrp gene regulatory elements, this mouse expresses tamoxifen-inducible CreERT2 recombinase. Additionally, the cassette includes a red fluorescent protein (tdTomato) for lineage tracing of Agrp-positive cells. In the absence of tamoxifen, CreERT2 recombinase remains cytoplasmic. Upon tamoxifen administration, CreERT2 translocates to the nucleus to mediate recombination. When Agrp-IRES-CreERT2-P2A-tdTomato mice are crossed with mice containing loxP sites, tamoxifen induction can trigger Cre recombinase-mediated sequence recombination between loxP sites in AgRP-positive neurons of the offspring.

The TGA stop codon was replaced with the P2A-CreERT2-WPRE-BGH pA cassette. CreERT2 recombinase is expressed under the regulatory control of Aqp5 gene elements. This model is a Tamoxifen-inducible Cre mouse, and when crossed with mice containing loxP sites, the offspring mice are expected to undergo sequence recombination between loxP sites mediated by Cre recombinase in alveolar type I cells, salivary gland alveolar cells, and gastrointestinal stem cells following Tamoxifen induction.