Non-coding RNAs are composed of modular secondary and tertiary structural units that themselves must form complex 3D structures for the entire RNA to fold and perform its biological function. However, the high density of negative charges associated with the phosphate backbone gives rise to substantial amounts of electrostatic repulsion, contributing to the large energetic barrier between unfolded and folded states. RNA folding chaperones (RFCs) are proteins that help lower this energy barrier by transiently interacting with RNA. Recent work has shown that some RFCs can function as macromolecular counterions for their RNA clients, thereby screening backbone electrostatic repulsion. However, the fundamental principles associated with how RFCs assist folding is largely unexplored. Here, we reduce RFCs to their smallest cationic constituents and then systematically “re-build” RFCs from these components (e.g., NH4+→ Lys→ polyK10→ RFC). We use single-molecule FRET to determine how these counterions influence both the relative population abundance and mean FRET efficiency of the folded and unfolded states of small model RNAs. From these studies, we have begun to determine aspects of RFCs that contribute to chaperoning RNA folding, such as net cationic charge and charge distribution. Here, we show that cationic amino acids (e.g., Lys and Arg) bind to RNA with apparent Kd values (appKd) on the order of 100 mM, which are comparable to those associated with their characteristic functional groups (e.g., ammonium and guanidinium). Further, increasing the net cationic charge of a counterion systematically decreases the appKd over several orders of magnitude. We also show a correlation between the decrease in appKd and a decrease in the folding free energy barrier. We then compare our observations of these polypeptide chaperones to the behavior of known RFCs (e.g., protamine, H1, and HCV nucleocapsid protein) with the small model RNAs.