Topologically protected magnetic skyrmions in magnetic materials are stabilized by interfacial or bulk Dzyaloshinskii-Moriya interaction (DMI). Interfacial DMI decays with increase of the magnetic layer thickness in just a few nanometers and bulk DMI only stabilizes magnetic skyrmions at low temperatures. Consequently, more flexibility in manipulation of DMI is required for utilizing nanoscale skyrmions in energy efficient memory and logic devices at room temperature (RT). Here, we demonstrate the observation of RT skyrmions stabilized by gradient DMI (g-DMI) in composition gradient engineered CoPt single layer films by employing topological Hall effect, magnetic force microscopy, and nitrogen vacancy scanning magnetometry. Skyrmions remain stable at a wide range of applied magnetic fields and are confirmed to be Bloch-type from micromagnetic simulation and analytical magnetization reconstruction. Furthermore, we observe skyrmion pairs which may be explained by skyrmion-antiskyrmion interactions. Our findings expand the family of magnetic materials hosting RT magnetic skyrmions by tuning g-DMI via gradient polarity and choice of magnetic elements.