The CD40LG gene is located on the X chromosome (Xq26.3) and encodes CD40 ligand (CD40L, also known as CD154), a type II transmembrane protein mainly expressed on activated T cells, platelets, and some B cells. Under inflammatory conditions, monocytes, natural killer cells, mast cells, and basophils can also be induced to express CD40L ^[1-3]. This protein binds to CD40 on the surface of antigen-presenting cells (such as B cells and dendritic cells), mediating key immune functions including T cell-dependent B cell activation, immunoglobulin class switching, and germinal center formation ^[3]. The CD40/CD40L interaction also regulates thrombosis, inflammatory responses, hematopoiesis, and the tumor immune microenvironment. Abnormal regulation of CD40LG is associated with X-linked hyper-IgM syndrome (XHIGM), a primary immunodeficiency disease characterized by recurrent infections due to defective antibody production ^[4]. In addition, abnormal expression of CD40L is involved in diseases such as systemic lupus erythematosus and atherosclerosis through its pro-inflammatory effects ^[5-6]. Due to the important role of the CD40/CD40L interaction in immune activation, CD40/CD40L has been an important target for immunotherapy. In recent years, significant progress has been made in CD40/CD40L-targeted therapy. Various drugs have been developed, including agonistic/antagonistic monoclonal antibodies, cellular vaccines, adenoviral vectors, and protein antagonists, and have shown therapeutic potential in malignant tumors, autoimmune diseases, and allograft rejection ^[7].
The B6-hCD40LG mice are a humanized model constructed by using gene editing technology to replace the endogenous extracellular domain of the mouse Cd40lg gene with the extracellular domain of the human CD40LG gene. This model can be used for research on the disease mechanisms and treatment methods of autoimmune diseases, cardiovascular diseases, cancers, etc., as well as for CD40LG-targeted drug development.

The STMN2 gene encodes the stathmin-2 protein, a microtubule-associated protein and member of the stathmin family. It plays a critical role in neuronal development, axonal growth, and regeneration by regulating microtubule dynamic stability. The STMN2 gene is predominantly expressed in the nervous system, with the highest levels in central and peripheral neurons, serving as a key molecule for maintaining axonal integrity and regenerative capacity. Additionally, its expression is elevated in developing neurons and is also present at moderate levels in adult brain tissues and the adrenal gland. Studies have shown that loss of TDP-43 function leads to cryptic splicing or premature polyadenylation of STMN2 mRNA, resulting in a significant reduction in functional STMN2 protein levels. This, in turn, causes neuromuscular junction denervation and axonal degeneration, which can be effectively reversed by exogenous supplementation of STMN2 or its post-translational stabilization ^[1-2]. In clinical research, the reduction of STMN2 due to TDP-43 pathology is a common feature in the majority of patients with amyotrophic lateral sclerosis (ALS) and can serve as a disease biomarker and potential therapeutic target ^[3]. Furthermore, STMN2 regulates α-synuclein (α-syn) aggregation and dopaminergic axonal integrity. It is significantly downregulated in the brain tissue of patients with Parkinson’s disease (PD), leading to dopaminergic neuron degeneration, elevated phosphorylated α-syn, and motor deficits. It has been identified as a key regulator functionally connected to known PD risk genes ^[4-5]. Studies have shown that humanized mouse models can precisely recapitulate STMN2-related pathological mechanisms. By delivering shRNA via adeno-associated virus (AAV), these models enable central nervous system-specific regulation, providing a robust tool platform for investigating the mechanisms of Parkinson’s disease (PD) ^[6]. The huSTMN2 mouse is a humanized model constructed using gene editing technology. The sequences from upstream of the exon 1 to downstream of the exon 5 of the mouse Stmn2 were replaced with the sequences from upstream of the exon 1 to downstream of the exon 5 of the human STMN2. The huSTMN2 mice can be used to investigate the pathogenesis and progression of neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS) and Parkinson’s disease (PD), facilitating the research and development of STMN2-targeted drugs and gene therapy strategies, as well as preclinical pharmacological and efficacy evaluations.

Agtr1b is located on chromosome 3 of mice. Nuclease Technology was used to design sgRNA; Agtr1b knockout mice were obtained by applying high-throughput electroporation of fertilized eggs. After sexual maturity, sperm were collected for cryopreservation.

Asic2 is located on chromosome 11 of mice. SgRNA and ssDNA were designed using Nuclease Technology; Asic2 conditional knockout mice were obtained by high-throughput electroporation of fertilized eggs. After sexual maturity, sperm were collected for cryopreservation.

Zfp959 is located on chromosome 17 of mice. Nuclease Technology will be used to design sgRNA; Zfp959 knockout mice will be obtained by applying high-throughput electroporation of fertilized eggs. After sexual maturity, sperm were collected for cryopreservation.

Zfp959 is located on chromosome 17 of mice. SgRNA and ssDNA will be designed using Nuclease Technology; Zfp959 conditional knockout mice will be obtained by high-throughput electroporation of fertilized eggs. After sexual maturity, sperm will be collected for cryopreservation.