Accurate and timely crop yield forecasts are critical for making informed agricultural policies and investments, as well as increasing market efficiency and stability. Earth observation data from space can contribute to agricultural monitoring, including crop yield assessment and forecasting. In this study, we present a new crop yield model based on the Difference Vegetation Index (DVI) extracted from Moderate Resolution Imaging Spectroradiometer (MODIS) data at 1 km resolution and the un-mixing of DVI at coarse resolution to a pure wheat signal (100% of wheat within the pixel). The model was applied to estimate the national and subnational winter wheat yield in the United States and Ukraine from 2001 to 2017. The model at the subnational level shows very good performance for both countries with a coefficient of determination higher than 0.7 and a root mean square error (RMSE) of lower than 0.6 t/ha (15–18%). At the national level for the United States (US) and Ukraine the model provides a strong coefficient of determination of 0.81 and 0.86, respectively, which demonstrates good performance at this scale. The model was also able to capture low winter wheat yields during years with extreme weather events, for example 2002 in US and 2003 in Ukraine. The RMSE of the model for the US at the national scale is 0.11 t/ha (3.7%) while for Ukraine it is 0.27 t/ha (8.4%).