The second-to-fourth digit ratio (2D:4D) is sexually dimorphic in humans, such that men on average have a lower 2D:4D than women. This somatic trait has been proposed as a biomarker for the organizational (permanent) effects of prenatal testosterone on the brain and behavior. Over the past few years, an accumulating research program has shown 2D:4D to be related to a multitude of sex-dependent, hormonally influenced psychological and behavioral traits. The present study investigated the 2D:4D ratio of 44 men and 70 women from 36 identical and 21 fraternal twin pairs. Both basic and advanced approaches for estimating heritability concordantly suggested that the trait is substantially heritable. The best-fitting structural equation model indicated that the contributions to individual differences in 2D:4D are 81% additively genetic, 19% nonshared environmental, and 0% shared environmental. Supplemental analyses showed that, consistent with a prediction from sex-hormone transfer theory, women from opposite-sex fraternal twin pairs had significantly lower (more male-typical) 2D:4D than women from same-sex fraternal twin pairs. Directions for research are discussed, such as investigating possible influences of the sex chromosomes on the expression of 2D:4D. Further family studies will be needed to test whether the transmission mode of 2D:4D is consistent with X-linked or Y-linked inheritance. The study of sex chromosome aberrations should indicate whether the presence of additional X or Y chromosomes is associated with 2D:4D levels.