Acteoside is an important bioactive natural product distributed in many plant species, composed of four moieties such as caffeic acid, glucose, rhamnose and phenylethyl alcohol, and possesses some bioactivities such as anti-inflammatory, anti-oxidant, neuro-protective, anti-tumor and so on. However, acteoside content in medicinal plants is low, and acteoside stability is bad, so acteoside biosynthesis is a problem. Recent years, acteoside biosynthesis pathway elucidation and bio-production have been widely investigated, so many achievements have been made up to now. In this study, we reviewed current advances in both the elucidation and bio-production such as the putative methods and enzymatic determination of acteoside biosynthesis pathway, functional analyses of the roles of some candidate genes for verbascoside biosynthesis by transgenic technology, acteoside production via metabolic engineering and synthetic biology approaches and plant tissue culture. Moreover, we first established a combined putative acteoside biosynthesis pathway based on its recent studies in animals, plants and microbes. Meanwhile, we pointed out both problems and shortcomings, and highlighted its future development trend. These results will provide references for the complete elucidation of acteoside biosynthesis pathway and the improvement of acteoside content in medicinal plants and acteoside production via microbial and plant metabolic engineering and synthetic biology approaches, and inform the readers critically of the latest developments of them.