Application of tumor necrosis factor α mutant zebrafish in Mycobacterium marinum infection
XIE Lingling1, WANG Fenghua2, WANG Decheng3, ZHAO Chao1, PENG Gang2, NIU Chen1
1. Key Laboratory of Medical Molecular Virology, Ministries of Education and Health, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China; 2. Institutes of Brain Science, Fudan University, Shanghai 200032, China; 3. Medical College of China Three Gorges University, Yichang 443002, China
Abstract：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.