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Rex rabbits have a unique fur structure with a variety of different coat colours. In this study, we analysed variability in the <em>agouti signalling protein</em> (<em>ASIP</em>) gene in Rex rabbits with different coat colours (castor, red, blue, chinchilla, otter and black). Three alleles at this locus were identified: A, light-bellied agouti (wild type), in castor and chinchilla Rex; <em>a</em>^t, black and tan, in otter, castor and chinchilla Rex; and <em>a</em>, black non-agouti, in black, red, blue, castor and chinchilla Rex rabbits. Two missense mutations, two synonymous substitutions and one indel were the identified polymorphisms associated to these three alleles, as already described by others. Gene expression of the <em>ASIP</em> gene was also evaluated in different tissues from animals of the six coat colours. <em>Agouti signalling protein</em> expression was always observed in all tissue/coat colour combinations.