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Plants are one of the vital sources of secondary metabolites. These secondary metabolites have diverse roles in human welfare, including therapeutic implication. Nevertheless, secondary metabolite yields obtained through the exploitation of natural plant populations is insufficient to meet the commercial demand due to their accumulation in low volumes. Besides, in-planta synthesis of these important metabolites is directly linked with the age and growing conditions of the plant. Such limitations have paved the way for the exploration of alternative production methodologies. Hairy root cultures, induced after the interaction of plants with Rhizobium rhizogenes (Agrobacterium rhizogenes), are a practical solution for producing valuable secondary metabolite at low cost and without the influence of seasonal, geographic or climatic variations. Hairy root cultures also offer the opportunity to get combined with other yield enhancements strategies (precursor feeding, elicitation and metabolic engineering) to further stimulate and/or enhance their production potential. Applications of metabolic engineering in exploiting hairy root cultures attracted the interest of several research groups as a means of yield enhancement. Currently, several engineering approaches like overexpression and silencing of pathway genes, and transcription factor overexpression are used to boost metabolite production, along with the contextual success of genome editing. This review attempts to cover metabolic engineering in hairy roots for the production of secondary metabolites, with a primary emphasis on alkaloids, and discusses prospects for taking this research forward to meet desired production demands. • Hairy roots (HRs) are an efficient system for secondary metabolites (SMs) production of therapeutic significance. • HRs provide various methods to modulate the SMs production including metabolic engineering. • HRs are shown to express pathway genes/transcription factors in single/multi-step engineering to boost desired SMs profile. • Gene silencing/genome editing in HRs significantly reduce the undesired metabolic routes to ensure desired SMs production. [ABSTRACT FROM AUTHOR]