This study identified gender-based constraints affecting the production, processing and marketing of biofortified cassava in two states in Nigeria, using a mixed methods approach. The study identified major differences between the two study sites (Benue and Oyo). The scale of production of biofortified cassava is higher in Oyo state among adult men because of their active involvement and collaboration with research institutes within the state and the ease of transporting products to Lagos State for designated diverse markets. However, in Benue state more adult and young women are engaged in cultivation, processing and marketing business to meet up with the increased demand due to higher consumer acceptance in this region. Gender analysis revealed that lack of access to hired-labour restricted the scale of production among women in especially Oyo state. Low product price and high price of processing equipment, poor market infrastructure and middle men exploitation were constraints significantly more mentioned by women in general. Majorly, the men identified limited processing facilities/equipment as the most important constraint affecting the demand of biofortified cassava roots, while generally women were more constrained by the shortage of basic amenities and trainings that hindered their processing efficiency. The study proposes integration of gender-responsive strategies to further enhance the delivery of biofortified cassava products in Nigeria.