Phenethylisoquinoline alkaloids (PIAs) are natural secondary metabolites with a tricyclic structure derived from l-tyrosine by a specific multienzymatic biosynthesized pathway. Colchicine is the most popular tricyclic proto-alkaloid of the phenethylisoquinoline group. It has garnered significant scientific interest due to its promising pharmaceutical applications in gout, cancer, and cardiovascular diseases. Colchicine and its derivatives have been found in limited plant species of the Colchicaceae family, such as Colchicum, Gloriosa, Merendera, Androcymbium, Sandersonia, etc. Despite significant progress made in understanding its biosynthesis and intermediate molecules through several radiotracer feeding, multiomics, and genetic engineering approaches, complete knowledge about its biosynthesis pathways and regulations is still not achieved. In vitro studies have shown that callus, cell suspension, and root cultures of C. autumnale and G. superba can produce colchicine in vitro, but the amount is very low compared to the demand. A comprehensive understanding of biosynthetic pathways with enzymological and genetic information could be a game changer in colchicine production by yield improvement in existing systems and by the establishment of new enhanced production systems through genetic and metabolic manipulations. This chapter provides a comprehensive overview of colchicine research, evaluating existing research on its biological sources, physiochemical properties, mode of action, toxicity profile, therapeutic potential, and biotechnological applications. It highlights gaps and discrepancies in the current understanding of colchicine research and is a foundation for future research. The chapter also discusses modern biotechnological options for fulfilling the increasing demand for pure colchicine, serving as a significant resource for researchers, students, and those interested in PIAs like colchicine.