Background: While biological effectiveness of PrEP in women who can adhere to daily dosage is strong, little is known about psychological and structural barriers to the PrEP program among transgender and non-transgender female sex workers in the U.S. Methods: Thirty semi-structured in-depth interviews and three focus group discussions among female sex workers were conducted in Washington D.C. between 2015 and 2016. All interviews were recorded, transcribed, and analyzed using NVIVO. Results: All study subjects were African American between 25 to 55 years old. Two-thirds were trans-gender women; more than half were local residents. Given the city's high HIV prevalence, overall perceived risk of HIV was high among female sex workers. Most HIV negative female sex workers were unaware of PrEP, but many had heard of Truvada as a treatment for HIV. Labeling effect, e.g. fear of being considered by peers and clients as HIV positive, was a primary concern for participating in PrEP; fear of drug-resistance and medicine's side effects followed in rank. Disclosure and the necessity to explain sex worker life style to physicians, fear of being discriminated against at clinics, concerns about cost and time spent at clinics, substance abuse, and low perceived health care need were also identified among potential barriers to PrEP. Discussions regarding harm reduction practices (condom use) during PrEP participation also were noted multiple times. Conclusion: Results showed high interest in PrEP among HIV negative transgender and non-transgender sex workers. Confidentiality, convenient locations, and client-centered health professionals or clinics may help attenuate participants' concerns.