This study explores gender differences in the roles of humor in the lives of Israeli children. Thirty-four Jewish middle-class Israeli children, sixteen girls and eighteen boys, aged between eight to ten years, were interviewed in focus groups in which they discussed a variety of humorous video segments, jokes, and everyday humor. The analysis suggests that humor in interaction is a highly gendered process in this age group and is employed differently by boys and girls to perform their gendered identities. Girls engaged much less in sexist and aggressive humor and clearly used it to maintain their separateness from boys and younger children. We conclude that humor provides us with another avenue through which to unveil the complicated processes of gender construction in pre-adolescent childhood, while demonstrating at the same time the ambivalence and complexity involved in these processes.