The anticancer anthracyclines doxorubicin and daunorubicin are used to treat a variety of cancers in dogs. The therapeutic utility of anthracyclines is limited by cardiotoxicity in some cases. Synthesis of anthracycline alcohol metabolites by carbonyl reductase 1 (CBR1) is crucial for the pathogenesis of cardiotoxicity. We hypothesize that genetic polymorphisms in canine cbr1 contribute to the variable pharmacodynamics of anthracyclines in dogs. DNA sequence variants in canine cbr1 were investigated in DNA samples from dogs of seven breeds. Thirteen SNPs were detected in canine cbr1. A 10-bp deletion in the 5'-untranslated region (5'-UTR) was found in specimens from the Labrador Retriever, Beagle, Siberian Husky, and Boxer breeds. The 5'-UTR also included a polymorphic "hot spot" region immediately downstream of the 10-bp deletion. DNA sequence variants in the "hot spot region" ranged from 1 to 21 bp in length. Bioinformatics searches identified a cluster of three to six potential binding sites for the transcription factor Sp1 in the DNA segment containing both the "hot spot" region and the 10-bp deletion. This information provides a foundation to allow us to investigate whether DNA sequence variants in the 5'-UTR of canine cbr1 impact the pharmacodynamics of anticancer anthracyclines in dogs.