Abstract The 11- cis-retinol dehydrogenase (11- cis-RoDH) gene encodes the short-chain alcohol dehydrogenase responsible for 11- cis-retinol oxidation in the visual cycle. The structure of the murine 11- cis-RoDH gene was used to reinvestigate its transcription pattern. An 11- cis-RoDH gene transcript was detected in several non-ocular tissues. The question regarding the substrate specificity of the enzyme was therefore addressed. Recombinant 11- cis-RoDH was found capable of oxidizing and reducing 9- cis-, 11- cis- and 13- cis-isomers of retinol and retinaldehyde, respectively. Dodecyl-β-1-maltoside used to solubilize the enzyme was found to affect the substrate specificity. This is the first report on a visual cycle enzyme also present in non-retinal ocular and non-ocular tissues. A possible role in addition to its role in the visual cycle is being discussed.