Transcription factors are key determinants that regulate cell fate decisions and tissue homeostasis. They also serve as vital mediators of extracellular signals. Transcription factors modulate gene expression typically by binding to specific sequences present in the regulatory region of target genes. As such, transcription factors are indispensable to translate cellular cues into a precise genomic response, leading to the activation or repression of gene transcription in a coordinated temporal, tissue-specific, and cell-specific manner. This is achieved through combinatorial interactions of different transcription factors, forming unique codes in each specialized cell type and in response to different physiological signals. Leydig cells are endocrine cells that, in response to various signals, produce two predominant hormones essential for male reproductive function: testosterone and INSL3. The synthesis of steroid hormones in Leydig cells requires the coordinated expression of several genes, a process that is highly reliant on transcription factors. This chapter aims to provide a comprehensive overview of the various transcription factors families identified as crucial regulators of Leydig cell gene expression and steroidogenesis.