肿瘤坏死因子α (tumor necrosis factor α，TNF-α)作为重要的炎性细胞因子，在类风湿关节炎、感染性休克等疾病的发病中起重要作用。为探究TNF-α在宿主抵抗海分枝杆菌感染中的作用，本研究采用成簇的规律间隔的短回文重复序列及其相关蛋白9〔clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9)，CRISPR/Cas9〕技术，使野生型斑马鱼TNF-α基因发生突变，成功构建并筛选出含有移码突变的TNF-α突变型(TNF-α^－/－)斑马鱼。利用原位杂交技术，发现TNF-α^－/－斑马鱼中TNF-α mRNA表达水平显著降低。与野生型斑马鱼相比，海分枝杆菌在TNF-α^－/－斑马鱼体内扩散和增殖更显著。结果提示，本研究成功构建TNF-α^－/－斑马鱼，有助于深入探究TNF-α在斑马鱼抵抗海分枝杆菌感染中的作用及其免疫调节机制。

Tumor necrosis factor α(TNF-α) is an important inflammatory cytokine implicated in the pathogenesis of rheumatoid arthritis, septic shock and other diseases. In order to investigate the functional role of host TNF-α in Mycobacterium marinum (M. marinum) infection, clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) (CRISPR/Cas9) technology was utilized to mutate the TNF-α gene in wild type (WT) zebrafish. A zebrafish containing a frame-shift mutation was successfully constructed, isolated, and named as TNF-α^－/－. It was found that the expression of TNF-α mRNA was significantly reduced in TNF-α^－/－ zebrafish by in situ hybridization (ISH). The proliferation of M. marinum in TNF-α^－/－ zebrafish was significantly elevated compared to WT zebrafish. The construction of TNF-α^－/－ zebrafish will allow further in-depth study on the function of TNF-α in mycobacterial infection and related immune regulation.