A new database of the Entomological Inoculation Rate (EIR) is used to directly link the risk of infectious mosquito bites to climate in Sub-Saharan Africa. Applying a statistical mixed model framework to high-quality monthly EIR measurements collected from field campaigns in Sub-Saharan Africa, we analyzed the impact of rainfall and temperature seasonality on EIR seasonality and determined important climate drivers of malaria seasonality across varied climate settings in the region. We observed that seasonal malaria transmission requires a temperature window of 15-40 degrees Celsius and is sustained if average temperature is well above the minimum or below the maximum temperature threshold. Our study also observed that monthly maximum rainfall for seasonal malaria transmission should not exceed 600 mm in west Central Africa, and 400 mm in the Sahel, Guinea Savannah and East Africa. Based on a multi-regression model approach, rainfall and temperature seasonality were significantly associated with malaria seasonality in most parts of Sub-Saharan Africa except in west Central Africa. However, areas characterized by significant elevations such as East Africa, topography has a significant influence on which climate variable is an important determinant of malaria seasonality. Malaria seasonality lags behind rainfall seasonality only at markedly seasonal rainfall areas such as Sahel and East Africa; elsewhere, malaria transmission is year-round. The study’s outcome is important for the improvement and validation of weather-driven dynamical malaria models that directly simulate EIR. It can contribute to the development of malaria models fit-for-purpose to support health decision-making towards malaria control or elimination in Sub-Saharan Africa.