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The exploration of functional materials relies greatly on the understanding of material structures and nanotechnologies. In the present work, chiral nematic cellulose nanocrystal (CNC) films were prepared by incorporation with four types of amino acids (AAs, glycine, histidine, phenylalanine, and serine) via evaporation-induced self-assembly. The films present ideal iridescence and birefringence that can be tuned by the amount of AAs added. The intercalation of AAs enlarged the pitch values, contributing to the red-shift trend of the reflective wavelength. Among the AAs, serine presented the most compatible intercalation into cellulose crystals. Interestingly, histidine and phenylalanine composite films showed high shielding capabilities of UV light in diverse wavelength regions, exhibiting multi-optical functions. The sustainable preparation of chiral nematic CNC films may provide new strategies for materials production from biocompatible lignocellulose.