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Melanoma differentiation-associated 7 (MDA-7) gene has also been classified as Interleukin 24 (IL-24), is a novel cytokine of the IL-10 family. The gene MDA-7/IL-24 possesses many unique features which make it an ideal target for cancer gene therapy. Genetic disorders that originated from single-gene mutations are the appropriate selection for gene therapy. On the other hand, cancer is a mutagenic disorder and by manipulating a single gene may not give desired and expected outcomes. In this regard, tumor suppressor gene (s) may give a promising therapeutic target for cancer treatment. Considering of MDA-7/IL-24 as an ideal therapeutic gene because it can selectively promote apoptosis in cancer cells without harming normal cells and able to encounter cancer on multiple levels. MDA-7/IL-24 has a classic role in the regulation of numerous signaling pathways, including activating biochemical changes in genetically diverse as cancer cells. Therefore, selectively targeting of MDA-7/IL-24 to tumor cells would provide enhanced efficacy and specificity. However, the role of MDA-7/IL-24 for cancer-specific gene therapy should be investigated in detail. MDA-7/IL-24 can affect multiple signaling pathways by activating biochemical changes specifically in genetically diverse as cancer cells. This chapter examines our current understanding of signaling pathways modulated by MDA-7, its unique features, current insights into the clinical trials of MDA-7/IL-24, and the problems/challenges associated with this gene.