Diffusion is essential for biochemical processes because it dominates molecular movement on small scales. Enzymatic reactions, for example, require fast exchange of substrate and product molecules in the local environment of the enzyme to ensure efficient turnover. On larger spatial scales, diffusion of secreted signaling proteins is thought to limit the spatial extent of tissue differentiation during embryonic development. While it is possible to measure diffusion in vivo, specifically interfering with diffusion processes and testing diffusion models directly remains challenging. The development of genetically encoded nanobodies that bind specific proteins has provided the opportunity to alter protein localization and reduce protein mobility. Here, we extend the nanobody toolbox with a membrane-tethered low-affinity diffusion regulator that can be used to tune the effective diffusivity of extracellular molecules over an order of magnitude in living embryos. This opens new avenues for future applications to functionally interfere with diffusion-dependent processes.