In current era of controlled release drug products and targeted drug delivery, niosomes are novel formulations of enormous significance. Niosomes provide greater formulation adaptability, more prominent physical cohesion and chemical perseverance. Niosomes can entrap lipophilic drugs and drugs of hydrophilic nature. Furthermore they augment the solubility of less soluble drugs in water, along with prolonged release effect. Cyclosporine A (CsA) is an immunosuppressant drug of exceptional importance, and is adopted as model drug. This drug has a low therapeutic index, and it has many toxic effects. After oral administration its bioavailability is variable due to poor absorption. So prime goal of this research was to formulate niosomal vesicles of cyclosporine A, to enhance its solubility and sustained release effect, consequently enhancing its bioavailability. Thin film hydration method was used for the preparation niosomes. Eleven niosomal formulations were successfully prepared. Nonionic surfactants and cholesterol was used in formulations.To determine the average drug content of CsA, HPLC method was used. This method was also validated as per protocols of ICH and used in determination of entrapment efficiency, in vitro and in vivo studies. The size range of niosomal formulations were from 415 nm to 1049 nm. The polydispersity index and zeta potential was in range of 0.259 to 0.572 and 23.8 to 35.2 mV respectively. Transmission electron microscopy revealed the spherical shape of niosomes in finally selected niosomal formulation F10. In the formulations F1, F2, F3, F4, F5 and F6 the formulation F2 exhibited the highest entrapment of 77.28%. In F2 the ratio of sorbitan monostearate and polysorbate 60 with cholesterol was 6:4. In formulations F7, F8, F9, F10 and F11, F10 achieved the maximum entrapment efficiency. In formulation F10 nonionic surfactants used were span 20 and brij 35 along with cholesterol in ratio (1:1). This formulation F10 exhibited maximum entrapment efficiency of 89.31%. No significant shift of peaks was found in ATR-FTIR analysis, which designates that there were no interactions.and CsA is compatible with niosomal components. DSC thermograms of niosomal formulations depicts that in niosomes the drug is amorphous in nature. Stability studies were conducted for three months, and it was found that niosomes were stable at 4°C and 25°C. But at refrigerated temperature 4-8 °C the amount of drug retained in niosomes was greater than at 25°C. In vitro drug release testing depicts improved dissolution along with controlled release behavior in all formulations (F 1 – F11).In vitro release studies at pH 1.2 and 7.4 showed that for all niosomal formulations the percentage release of drug was significantly greater as compared to drug aqueous dispersion. Formulation F10 having span 20 and brij 35 mixed surfactants, presented better dissolution and augmented sustain release rates in comparison with other formulations. Kinetic modeling of drug release of niosomal vesicles exhibited that they follow zero order release. To calculate the release exponent (n) kinetic model of korsemeyer and peppas was used. It exhibited that drug transport mechanism is anomalous. For in-vivo studies selected optimized niosomal formulations F2 and F10 were figure out. As a control CsA aqueous dispersion was used. PK Solver software was used for analysis of pharmacokinetic parameters. The values of AUC 0-inf, Tmax and MRT 0-inf of niosomal formulations F10 and F2 were significantly greater than the CAD, confirming the sustained release and improved bioavailability of CsA. However, F10 formulation displayed greater AUC 0-t, Cmax and mean residence time as compared to F2, due to mixed surfactant system of span 20 and brij 35 used in F10 formulation along with cholesterol which improves the bioavailability and results in more sustain release effect of CsA. So it was found that niosomal formulations based on mixed surfactants, is effective delivery system for prolonged delivery of CsA along with improved oral bioavailability.