Cells employ signaling pathways to make decisions in response to changes in their immediate environment. Transforming growth factor beta (TGF-beta) is an important growth factor that regulates many cellular functions in development and disease. Although the molecular mechanisms of TGF-beta signaling have been well studied, our understanding of this pathway is limited by the lack of tools that allow the control of TGF-beta signaling with high spatiotemporal resolution. Here, we developed an optogenetic system (optoTGFBRs) that enables the precise control of TGF-beta signaling in time and space. Using the optoTGFBRs system, we show that TGF-beta signaling can be selectively and sequentially activated in single cells through the modulation of the pattern of light stimulations. By simultaneously monitoring the subcellular localization of TGF-beta receptor and Smad2 proteins, we characterized the dynamics of TGF-beta signaling in response to different patterns of blue light stimulations. The spatial and temporal precision of light control will make the optoTGFBRs system as a powerful tool for quantitative analyses of TGF-beta signaling at the single cell level.