Targeted gene replacement (TGR) is an important technique for gene function analysis. With the development of genome sequencing and transformation, TGR has been applied widely to filamentous fungi, and many new systems and approaches have been established. In this paper, strategies involved in TGR in filamentous fungi were reviewed, including transformation systems, targeting vector construction, mutant selection and so on. Compared with TGR, RNAi and other techniques of gene function analysis were introduced and reviewed as well.