Introduction: The Transgender Awareness Webinar was designed to educate the general public about transgender individuals and reduce stigmatizing attitudes toward transgender and gender diverse people. Methods: The present study involved the development of an update of the webinar to be more inclusive of non-binary gender identities and to evaluate the revised webinar using a pilot randomized control trial (RCT) design. The study included 203 participants (n = 102 treatment group; n = 101 control group). Each participant completed a pre-test and post-test of the Transphobia Scale before and after participation in the webinar. Results: Results indicated that transphobia scores went down from pre-to-posttest among participants in the high baseline transphobia category. In addition, several demographic groups had significantly higher levels of mean transphobia scores (higher income, male gender, traditional religious groups). Conclusions: The results of this study suggest that the updated Transgender Awareness Webinar holds promise for reducing transphobia among members of the general public who have high levels of baseline transphobia. Other findings and implications are presented with regard to the development of an anti-stigma webinar.