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Proniosomes refer to a flexible vesicular carrier with the potential for drug administration through the transdermal route. A proniosome gel type transdermal delivery system of Atenolol was prepared and extensively studied both in vitro drug release and ex vivo permeation studies. The prepared formulations were evaluated for vesicle size, entrapment efficiency, in vitro drug loading, and drug release studies. The release of drug had shown considerable improvement in controlled manner from the prepared gel formulation. It was observed that Span 40 & 60 (A 8) based formulations shows vesicles of minimum size and higher entrapment efficiency compared to the other formulations. Proniosomal transdermal therapeutic system (A 8) was found to be the optimized formulation as it possess good drug release and shows permeation in a steady-state manner over a desired period of time. Also the drug diffusion across snake sheded skin, guinea pig abdomen skin, albino rat, porcine ear correlates better with in vitro drug release studies. The formulation was found to be stable when stored at room temperature and at refrigeration temperature (4 ± 2°C) for 90 days.