Transgender individuals face significant healthcare disparities, often exacerbated by provider prejudice and knowledge gaps. Nurses, as frontline providers, play a critical role in promoting inclusive care. This study aimed to evaluate the levels and predictors of transphobia and attitudes toward transgender individuals among nurses in Türkiye. : A cross-sectional survey was conducted with 100 registered nurses. Participants completed the Transphobia Scale, the Attitudes Toward Transgender Individuals Scale, and the Hudson & Ricketts Homophobia Scale. Additional measures included perceived knowledge, prior education on transgender health, familial attitudes, and comfort levels when working with transgender patients. Multiple linear regression analyses identified predictors of transphobia and attitudes. : The mean Transphobia Scale score was 41.53 (SD = 12.67), and the mean Attitudes Toward Transgender Individuals Scale score was 57.45 (SD = 16.42). Greater homophobia, negative parental attitudes, and higher perceived knowledge significantly predicted higher transphobia scores (Adjusted R= 0.327). Prior education on transgender health was also associated with lower transphobia. Lower comfort with transgender patients was marginally associated with higher transphobia. More positive attitudes toward transgender individuals were predicted by lower transphobia, lower homophobia, and prior education (Adjusted R = 0.526). : Findings underscore the urgent need for structured transgender health education and culturally sensitive interventions among nurses. Addressing sociocultural factors and training gaps could enhance equitable healthcare delivery for transgender populations, particularly in culturally conservative settings like Türkiye.