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Increasing plants' photosynthetic efficiency is a major challenge that must be addressed in order to cover the food demands of the growing population in the changing climate. Photosynthesis is greatly limited at the initial carboxylation reaction, where CO₂ is converted to the organic acid 3-PGA, catalyzed by the RuBisCO enzyme. RuBisCO has poor affinity for CO₂, but also the CO₂ concentration at the RuBisCO site is limited by the diffusion of atmospheric CO₂ through the various leaf compartments to the reaction site. Beyond genetic engineering, nanotechnology can offer a materials-based approach for enhancing photosynthesis, and yet, it has mostly been explored for the light-dependent reactions. In this work, we developed polyethyleneimine-based nanoparticles for enhancing the carboxylation reaction. We demonstrate that the nanoparticles can capture CO₂ in the form of bicarbonate and increase the CO₂ that reacts with the RuBisCO enzyme, enhancing the 3-PGA production in in vitro assays by 20%. The nanoparticles can be introduced to the plant via leaf infiltration and, because of the functionalization with chitosan oligomers, they do not induce any toxic effect to the plant. In the leaves, the nanoparticles localize in the apoplastic space but also spontaneously reach the chloroplasts where photosynthetic activity takes place. Their CO₂ loading-dependent fluorescence verifies that, in vivo, they maintain their ability to capture CO₂ and can be therefore reloaded with atmospheric CO₂ while in planta. Our results contribute to the development of a nanomaterials-based CO₂-concentrating mechanism in plants that can potentially increase photosynthetic efficiency and overall plants' CO₂ storage.