The skeleton supports body structures in vertebrates and helps maintain calcium homeostasis throughout life. Disruption of genes involved in mammalian bone formation has often led to embryonic lethality, hence preventing study of these genes’ role in adult animals. To develop a usable tool for such study, we generated transgenic mice in which a 2.3-kb mouse Col1a1 proximal promoter, which is active in all osteoblasts, drives a transgene coding for a polypeptide consisting of Cre recombinase fused to a mutated ligand-binding domain of the estrogen receptor. In this Col1a1-CreERT2 mouse line, expression patterns of the transgene and of the resulting Cre-mediated DNA recombination are analyzed by crossing with ROSA26 reporter mice and by measurement of β-galactosidase activity and X-gal staining. Exposure to 4-hydroxytamoxifen induced Cre-mediated recombination in osteoblasts in virtually all bones and in odontoblasts in teeth of both embryos and postnatal mice. The generation of these transgenic mice provides a new and important tool with which to study the function of specific genes in bone and tooth physiology and diseases in intact animals after birth.