This dataset represents results from this study attributed to the NHDPlus V2 catchments. Changes in climate occurring throughout the Mississippi River Basin are expected to lead to additional impacts in stream habitats and fish assemblages in multiple ways, including changing changing thermal properties and flow regimes. To manage streams for current impacts and future changes, managers need region-wide information for decision-making and developing proactive management strategies. Our project provides a suite of climate metrics that have been found to be relevant to the distribution and population structure of aquatic organisms in freshwater stream networks. These results provide natural resource managers, decision-makers, and the public with a suite of climate metrics summarized for all catchments within the Mississippi River Basin. This data set is comparable to our previous research conducted across the Northeast Climate Science Center region which is also located in ScienceBase ( https://doi.org/10.5066/F7GQ6W7C).