Abstract: Taxol (generic name paclitaxel) is a highly effective anti-cancer agent widely used in the clinical treatment of various cancers. At present, the production of paclitaxel is primarily based on artificial semi-synthesis from extracted intermediates (such as baccatin Ⅲ). This commercial method still relies on Taxus plant resources, resulting in high medical costs. The rise of synthetic biology provides a novel approach to acquire sufficient paclitaxel, but research on its biosynthesis is yet to be advanced. Recently, multi-omics approaches have been applied in biosynthesis research on paclitaxel. In the present paper, we review recent progress on the biosynthesis, regulation, and heterologous production of paclitaxel aided by multi-omics, providing new insights into paclitaxel biosynthesis.