Filamentous fungi are important industrial microorganisms. The focus on its metabolic engineering is to optimize the metabolic pathway with gene expression regulation technology to meet with the industrial production needs. Antisense RNA technology due to its simplicity compared with the gene knock-out technology has great perspectives in filamentous fungal metabolic control. It is an efficient method for regulating gene expression and a key tool for metabolic engineering. In this article, we addressed the mechanism of antisense RNA technology and its applications in filamentous fungal metabolic engineering. Additionally, future perspectives were discussed.