The domestic cat (Felis silvestris catus) is quickly becoming the most popular animal companion in the world. The evolutionary processes that occur during domestication are known to have wide effects on the morphology, behaviour, cognition and communicative abilities of a species. Since facial expression is central to human communication, it is possible that cat facial expression has been subject to selection during domestication. Standardised measurement techniques to study cat facial expression are, however, currently lacking. Here, as a first step to enable cat facial expression to be studied in an anatomically based and objective way, CatFACS (Cat Facial Action Coding System) was developed. Fifteen individual facial movements (Action Units), six miscellaneous movements (Action Descriptors) and seven Ear Action Descriptors were identified in the domestic cat. CatFACS was then applied to investigate the impact of cat facial expression on human preferences in an adoption shelter setting. Rehoming speed from cat shelters was used as a proxy for human selective pressure. The behaviour of 106 cats ready for adoption in three different shelters was recorded during a standardised encounter with an experimenter. This experimental setup aimed to mimic the first encounter of a cat with a potential adopter, i.e. an unfamiliar human. Each video was coded for proximity to the experimenter, body movements, tail movements and face movements. Cat facial movements were not related to rehoming speed, suggesting that cat facial expression may not have undergone significant selection. In contrast, rubbing frequency was positively related to rehoming speed. The findings suggest that humans are more influenced by overt prosocial behaviours than subtle facial expression in domestic cats.