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Sport is an agent of social change, but that change does not always track in a progressive direction. Sport can be a site for contesting and reversing the gains of progressive social movements as much as furthering the values of equality and justice for historically marginalized groups. This dynamic of contestation and reversal is now playing out in a new wave of anti-transgender backlash that has gained adherents among some proponents of equal athletic opportunities for girls and women. In this latest twist in the debate over who deserves the opportunity to compete, the sex-separate athletic programming permitted by Title IX has been the vehicle for depicting trans athletes – and especially trans girls – as unwelcome intruders poised to take away Title IX’s gains for female athletes. The trans threat narrative relies on and reinforces a dichotomy between trans girls and cisgender girls, with the latter positioned in this narrative as the “real” girls, and the former depicted as suspect subjects – and even, absurdly, as boys posing as girls for opportunistic reasons. Despite the lack of empirical evidence that trans athletes pose any threat to girls in sports, the threat narrative has been an effective strategy for reinscribing traditional understandings of sex and gender roles precisely because it trades on the popularity of Title IX across the political spectrum. The trans threat narrative has even gained adherents among some advocates for girls in sports who otherwise align with liberal supporters of transgender rights. The threat narrative has also succeeded in gaining support among state legislators, school boards, and parents, under the banner of protecting sports opportunities for “girls.” The issue has divided the women’s sports community, which has historically functioned as a unified front for advancing gender equality in athletics. This article examines the threat narrative in relation to the theories and justifications for Title IX’s baseline of sex-separation in school sports programs. It contends that the narrative embraces the most problematic of the justifications for sex-separation of sports, thereby reinforcing stereotypes of gender difference that have long thwarted girls’ and women’s efforts to achieve equal athletic opportunity. Equality for girls and women in sport is best achieved by embracing trans inclusion and rejecting efforts to exclude trans athletes from competition.