In Canada, indigenous women are overrepresented among new HIV infections and street-based sex workers. Scholars suggest that Aboriginal women’s HIV risk stems from intergenerational effects of colonisation and racial policies. This research examined generational sex work involvement among Aboriginal and non-Aboriginal women and the effect on risk for HIV acquisition. The sample included 225 women in street-based sex work and enrolled in a community-based prospective cohort, in partnership with local sex work and Aboriginal community partners. Bivariate and multivariate logistic regression modeled an independent relationship between Aboriginal ancestry and generational sex work; and the impact of generational sex work on HIV infection among Aboriginal sex workers. Aboriginal women (48%) were more likely to be HIV-positive, with 34% living with HIV compared to 24% non-Aboriginal. In multivariate logistic regression model, Aboriginal women remained 3 times more likely to experience generational sex work (aOR:2.97; 95%CI:1.5,5.8). Generational sex work was significantly associated with HIV (aOR=3.01, 95%CI: 1.67–4.58) in a confounder model restricted to Aboriginal women. High prevalence of generational sex work among Aboriginal women and 3-fold increased risk for HIV infection are concerning. Policy reforms and community-based, culturally safe and trauma informed HIV prevention initiatives are required for Indigenous sex workers.