The following is a purely theoretical experimental design for academic discussion only and does not constitute any guidance for direct implementation on humans. Before actually conducting such research, rigorous scientific demonstration, ethical review, regulatory approval, and long-term safety verification must be carried out. Please be sure to follow the relevant international and national regulations to ensure that all research is carried out under the premise of strict control and respect for the rights and safety of the subjects. [Overall idea] The goal is to change or regulate key molecules related to innate/adaptive immunity (such as factors that regulate antiviral response, inflammatory response, and cytokine secretion) through gene editing, so that the body can produce a faster, stronger, and more appropriate immune response when encountering pathogens, reducing the risk of infection. Based on the maturity of gene editing technologies such as CRISPR/Cas9, CRISPRa/CRISPRi, it is recommended to verify the strategy in in vitro models and animal models before considering clinical transformation. [Steps Overview] 1. Screening and verification of target genes a. Literature research and bioinformatics analysis: Select genes closely related to anti-disease immunity, such as genes related to interferon pathways, T cell activation, and regulation of inflammatory responses (such as IRF, STAT, and cell surface receptors, etc.). b. Functional validation: Use in vitro cell models (such as human immune cell lines or immune cells differentiated from induced pluripotent stem cells (iPSCs)) to study the correlation between target gene expression and cell disease resistance. RNA interference, overexpression and other methods can be used to preliminarily verify the effect of the change on the immune response. 2. Design gene editing strategy a. Editing scheme selection: – If the goal is to enhance the expression of an antiviral or antibacterial factor, CRISPR activation technology (CRISPRa) can be used to increase the endogenous expression of the gene; – If the goal is to eliminate negative immune regulatory factors, CRISPR/Cas9 can be used to knock out or knock down related genes. b. Editing element construction: Design specific sgRNA and construct the corresponding Cas9 expression vector or dCas9-based regulatory system. Perform in vitro scoring to ensure that off-target effects are within controllable range. 3. In vitro experimental stage a. Cell model selection: Use human immune cell lines (such as T cells, macrophages or dendritic cells) or primary immune cells differentiated from iPSCs. b. Gene editing operation: Introduce editing tools into cells through conventional methods such as electroporation and viral vector transfection. c. Verify the editing effect: – Molecular level: Use PCR, Sanger sequencing, and deep sequencing to detect editing accuracy and off-target conditions; – Functional level: Confirm the increase or knockout of target gene expression through mRNA and protein detection (qPCR, Western blot). d. Functional test: – Immune response detection: Stimulate cells with pathogen-related molecules (such as viral components, bacterial cell wall fragments, etc.) to detect the secretion levels of interferon, inflammatory factors, etc.; – Pathogen challenge: Under strict biosafety conditions, use simulated infection to detect the response and killing ability of edited cells to pathogen invasion. 4. Animal model verification stage a. Construct transgenic animals: – First, gene editing can be used in mouse models (or using established humanized mouse models) to introduce corresponding genetic modifications. – Design a control group (wild type) and an experimental group (edited type), and ensure that the sample size is sufficient for statistical analysis. b. Functional evaluation: – Evaluate routine indicators of the immune system (immune cell subsets, cytokine expression, etc.) in edited animals; – Conduct challenge tests with appropriate pathogens (viruses, bacteria or parasites) to observe infection rates, pathological changes and survival rates. c. Safety evaluation: – Check for abnormal inflammation, immune hyperreaction or other systemic side effects; – Long-term follow-up to evaluate whether immune-related autoreactions or other pathological changes occur. 5. Turn to preclinical research and ethical review a. Non-human primate experiments: After the encouraging results of mouse experiments, the effectiveness and safety of the editing strategy can be further verified in non-human primates. b. Formulate clinical trial plans: Based on sufficient preliminary data, after obtaining approval from the ethics committee and regulatory agencies (such as the FDA or the corresponding agencies of the country), strictly controlled Phase I clinical trials (mainly focusing on safety) can be initiated. c. Screening of the test population, informed consent and long-term follow-up: All steps must comply with ethical standards and relevant laws and regulations. [Precautions and risk warnings] 1. Gene editing technology still has off-target risks, which may cause unknown adverse consequences. 2. Changing the balance of the immune system may cause autoimmune diseases, chronic inflammation or excessive immune response. 3. The editing of the human genome involves major ethical and legal issues. Currently, germline cell editing is strictly prohibited worldwide, and somatic cell editing must also be extremely cautious. 4. Any clinical research intended for human use must undergo a multi-level, rigorous ethical and scientific review process. [Summary] This plan proposes an experimental design idea from gene screening, in vitro validation to animal model validation, to preclinical research and ethical review, with the aim of theoretically improving the immune system's resistance to pathogens through gene editing. It should be emphasized that the technical, ethical and safety issues involved in actual operations are extremely complex. All work must be carried out under the supervision of multidisciplinary experts, ethics committees and regulatory agencies. Do not try it yourself or simply promote it. Reminder again: The above content is only for academic theoretical discussion and should not be used to guide any actual human gene editing experiments that have not been strictly reviewed.