Exosome-like Particles (ELPs) derived from natural products have already attained great interest in drug delivery and therapeutic applications. Among these, mushroom-derived exosome-like particles (MELPs) remain an unexplored group with immense biomedical potential. Since the medicinal and edible mushrooms are rich in bioactive compounds such as polysaccharides, proteins, and phenolic acids, there is a great chance that their ELPs also carry these compounds during their biogenesis. Hence, MELPs with these beneficial compounds, exhibiting favorable conditions such as nano-size, stability, and cellular uptake properties, should be considered a novel potential natural nano-carrier. Here, we detail the current understanding of MELPs in terms of their biological functions, physicochemical characteristics, molecular cargo, and isolation methods. We also highlight their potential applications in cancer therapy, immune modulation, inflammation, and microbiome regulation. Moreover, we outline the challenges and future directions for standardizing the isolation protocol to yield large-scale production and elucidate the mechanistic pathways. As MELP’s research advances, it may emerge as the next generation of nano-carriers in natural therapeutics and green nanomedicine.