Objective We aimed to establish a novel resource consisting of genetically diverse mice and perform a preliminary characterization of the genetic traits of these mice. Methods (1) A multi-parent panel was used to create the Genetic Diversity mice through appropriate crosses. ( 2 ) Whole-exon sequencing was used to analyze the genetic characteristics of the established Genetic Diversity outbred and inbred mice. ( 3) Using the genome sequences of the founders and offspring, we created the chromosome physical maps of these mice and calculated the founder contributions. Results (1) Genetic Diversity inbred mice were characterized by stable inheritance and consistent phenotype and were suitable for rapid phenotypic correlation analysis. (2) Compared with the inbred mice, the Genetic Diversity outbred mice showed superior growth and reproductive performance, and more genetic variation, which led to a “ mosaic-like ” chromosomal pattern. As unique individuals, the outbred mice are ideal for the study of subtle genetic regulatory mechanisms. (3) The founder contributions to the progeny lines were not uniform and genetic preference existed among the offspring. Conclusions Compared with the more conventional inbred mice, the Genetic Diversity mice established by breeding from multiple lines possessed diverse genetic variations. The Genetic Diversity outbred and inbred lines are associated with pros and cons but complement one another, and together represent a novel and powerful tool for the study of complex genetic traits.