This study was intended to develop and optimize the transethosomal gel of Bifonazole for the treatment of fungal infections. For the optimization purpose, Box-Behnken Design was used, and the concentration of soya lecithin, ethanol and sodium cholate were selected as independent variables. The individual and combined effects of independent variables were assessed on vesicle size, PDI and entrapment efficiency. Further, the optimized Bifonazole-loaded transethosomes were evaluated for zeta potential and TEM. The optimized transethosomes were incorporated into 1.5% Carbopol 940 gel and evaluated for in vitro release, ex vivo permeation studies and antifungal activity. The vesicle size, PDI, %EE and zeta potential of optimized formulation were found to be 104.7nm, 0.356, 86.23%, and -27.2mV respectively. The transmission electron microscopy showed that the vesicles were uniform and spherical. The in vitro release of transethosome suspension (73.49%) and transethosomal gel (66.43%) was more when compared to the conventional gel (45.37%), marketed cream (43.31%), and drug suspension (36.49%). For both prepared gel in vitro release data fits well into the Higuchi's model (R2 > 0.99). From an ex vivo study, transethosomal gel shows a significant increase in the steady-state flux to 1.53 times than the conventional gel and 1.54 times than marketed cream. The prepared transethosomal gel of Bifonazole showed better antifungal activity than the marked cream. This study concluded that Bifonazole-loaded transethosomal gel showed better permeation and enhanced antifungal activity.