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The relationship between protein N-myristoylation and innate immunity has emerged as an exciting area of research. Here, the recent progress of protein N-myristoylation and modulation in innate immune responses, including TLR4 inflammatory responses, demyristoylation-induced innate immunosuppression during Shigella flexneri infection, and human immunodeficiency virus-1 (HIV-1) assembly, intracellular host interactions and spread, is summarized.