Atopic dermatitis (AD) is a common chronic skin condition characterised by recurring inflamed and itchy skin. The actin remodelling protein, Flightless I (Flii) can regulate tissue inflammation via its effects on Toll-like Receptor (TLR) signalling. Here we investigated if changing the expression of Flii in an in vivo mouse model of AD affects the development and severity of this inflammation-mediated skin condition. Using Flii heterozygous (Flii+/-), wild-type (Flii+/+) and Flii transgenic (FliiTg/Tg) mice in an ovalbumin murine model of AD, we showed that mice with reduced levels of Flii have less severe dermatitis, characterised by decreased erythema and reduced transepidermal water loss compared to wild-type and Flii-overexpressing mice. In contrast Flii-overexpressing mice showed increased erythema. Histological examination of these mice with ovalbumin-induced dermatitis further showed that Flii-deficient mice had decreased skin thickness and dermal cellular infiltrate compared to wild-type and Flii-overexpressing mice. Using toluidine blue staining, mast cells numbers were also observed to be reduced in the Flii-deficient mice. The results suggest that reducing Flii levels in the skin dampens the symptoms associated with AD. Further studies will be performed to elucidate the mechanisms involved including the relationships with TLRs.