The lesbian, gay, bisexual, and transgender (LGBT) community continues to experience worse health outcomes than their heterosexual counterparts. Inequities in health care include low health insurance rates, high rates of stress due to systemic discrimination and stigma, and a lack of cultural competency in the health care system. Gender and sexual minority (GSM) people are at higher risk of mental health disorders, sexually transmitted infections (STIs) and human immunodeficiency virus (HIV), substance use and abuse, cancer, suicide, and other disorders/diseases. A lack of cultural competency in health care systems perpetuates these health disparities and inequities in care that burden the LGBT community. This project’s purpose was to evaluate the effectiveness of an online educational intervention on enhancing health professionals’ cultural competence of GSM health. This study used a one-group pre-, post-, and follow-up survey intervention, quasi-experimental design to evaluate the effectiveness of an educational intervention on improving health professionals’ knowledge, clinical preparedness, and attitudinal awareness of GSM health. The study’s setting was at a primary care center with clinics spread across three rural counties in North Dakota with a combined population of less than 14,000. This study used convenience sampling, and the recruitment of participants included a project presentation at the health care organization where the project would take place and an email invitation. Thirty-six participants completed the pre-survey, 11 of those 36 participants completed the educational intervention and post-survey, and six of those 11 participants completed the follow-up survey four weeks after completing the educational intervention. The instrument used for the presurvey, post-survey, and follow-up survey was the Lesbian, Gay, Bisexual, and Transgender Development of Clinical Skills Scale (LGBT-DOCSS). A paired sample t-test was used to compare pre, post, and four-week follow-up LGBT-DOCSS mean scores. The results indicated a statistically significant improvement in LGBT-DOCSS mean score on the post-survey (p = 0.0011) and four-week follow-up (p = 0.01) compared to the pre-survey. Additionally, the majority of participants reported that this educational intervention was valuable to their practice. This project revealed that an online educational intervention effectively enhanced health professionals’ cultural competence of GSM health.