Abstract Objective The aim of this study was to determine if FAM83H over-expression causes dentine or enamel malformations. Materials and methods The full-length mouse Fam83h cDNA was inserted into the pCAGIG vector between a β-actin promoter and β-globin enhancer for ubiquitous expression in transgenic mice. Recombinant mouse FAM83H was expressed and used to generate polyclonal antibodies. Western blots showed enhanced expression of the Fam83h transgene. The effects of transgene expression on tooth development were assessed by microhardness measurements of enamel and dentine. Total thickness of incisor enamel at the level of the alveolar crest was measured and decussating rod patterns were visualized by scanning electron microscopy (SEM). Results Three transgenic mouse lines were selected based upon their transgene expression levels. There was no statistically significant difference in the Vickers microhardness values of enamel or dentine between the transgenic lines or between the transgenic lines and wild type mice. No statistically significant differences in enamel thickness were observed between the transgenic lines and the wild type mice. SEM analysis revealed no apparent differences in the enamel crystal and rod morphologies. Conclusion Our findings demonstrate that over-expression of FAM83H in mice does not produce a phenotype in dentine or enamel.