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Ageritin is a specific ribonuclease, extracted from the edible mushroom <i>Cyclocybe aegerita</i> (synonym <i>Agrocybe aegerita</i>), which cleaves a single phosphodiester bond located within the universally conserved alpha-sarcin loop (SRL) of 23–28S rRNAs. This cleavage leads to the inhibition of protein biosynthesis, followed by cellular death through apoptosis. The structural and enzymatic properties show that Ageritin is the prototype of a novel specific ribonucleases family named ‘ribotoxin-like proteins’, recently found in fruiting bodies of other edible basidiomycetes mushrooms (e.g., Ostreatin from <i>Pleurotus ostreatus</i>, Edulitins from <i>Boletus edulis</i>, and Gambositin from <i>Calocybe gambosa</i>). Although the putative role of this toxin, present in high amount in fruiting body (>2.5 mg per 100 g) of <i>C. aegerita,</i> is unknown, its antifungal and insecticidal actions strongly support a role in defense mechanisms. Thus, in this review, we focus on structural, biological, antipathogenic, and enzymatic characteristics of this ribotoxin-like protein. We also highlight its biological relevance and potential biotechnological applications in agriculture as a bio-pesticide and in biomedicine as a therapeutic and diagnostic agent.