Trembling Before G-d interviews and follows several gay and lesbian Orthodox Jews, many only seen in silhouette, and also interviews several rabbis and psychologists regarding their views on homosexuality in Orthodox Judaism. The film repeatedly returns to several characters:
David is an observant Orthodox Jewish doctor from Los Angeles who has spent a decade trying to reconcile his homosexuality with Judaism. He has tried numerous forms of "treatment", from eating figs and praying to wearing a rubber band on his wrist to flick whenever he thinks of men, but to no avail. During the course of the film, David decides to visit the Chabad rabbi to whom he first came out.
Israel David Fishman is a 58-year-old New Yorker who decided he could not be gay and Orthodox, and turned his back on his religion, though not before his family forced him into electroshock therapy to try to cure him. Now a tour guide in the Haredi neighborhoods of New York, the film follows him as he gives a tour, psychoanalyzes himself and decides, on the 25th anniversary of being with his life partner, to call his 98-year-old father, a rabbi, whom he has not seen in over twenty years.
Michelle is another New Yorker, in her forties, who believed she was the only Hasidic lesbian in the world and as a consequence allowed herself to be pressured into marriage. However, she got divorced and was subsequently ostracized by her family and community when they discovered she was homosexual. The film shows her visiting her old neighborhood and an Orthodox fair.
Rabbi Steven Greenberg, one of the founding members of the Jerusalem Open House, a gay rights organization in Israel which provides support to gay Orthodox Jews and their families, who is sometimes called "the world’s first openly gay Orthodox rabbi", discusses parents' reactions to their children coming out, as well as traditional interpretations of the prohibitions on homosexual acts in the Torah.
Shlomo Ashkenazy is a gay psychotherapist who has run a confidential support group for Orthodox gay men for nearly 20 years. He is interviewed about the effects of Orthodox attitudes to homosexuality and the reactions of rabbis to gay Jews.
Mark is the English son of a Haredi rabbi. Coming out at 15, he was expelled from seven yeshivas for homosexual activity before becoming a drag queen, and is now dying of AIDS-related illness. He visits several yeshivas and other religious sites throughout the film. He remains upbeat, at one point saying, "Being a Jew is such a nice present to receive."