An AIEgen-Based Carrier Enables Efficient Cytosolic Delivery of Bioactive Proteins

Yu Yang,1– 3,* Huixia Jia,1– 3,* Lin Yang,4 Benzhao He,4 Ke Zhang,1– 3 He Liu1– 3 1Department of Systems Science, Faculty of Arts and Sciences, Beijing Normal University, Zhuhai, Guangdong, 519087, People’s Republic of China; 2International Academic Center of Complex Systems, Beijing Normal University, Zhuhai, Guangdong, 519087, People’s Republic of China; 3School of Systems Science, Beijing Normal University, Beijing, 100875, People’s Republic of China; 4Department of Chemistry, Faculty of Arts and Sciences, Center for Advanced Materials Research, Beijing Normal University, Zhuhai, Guangdong, 519087, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ke Zhang, Email kezhang@bnu.edu.cn He Liu, Email heliu@bnu.edu.cnBackground: Cytosolic protein delivery enables the direct transport of exogenous proteins into the cytoplasm, offering a powerful approach for precise regulation of intracellular activities. This strategy facilitates the investigation of complex physiological processes in cellular and molecular biology and supports the development of protein-based biotechnologies and precision therapeutics, such as targeted protein supplementation for cellular repair or modulation of aberrant cellular functions. However, current delivery methods face some challenges, including cytotoxicity, immune activation, and the difficulty of maintaining protein stability and activity during transport.Methods: Here, we report a novel AIEgen (Aggregation-induced emission luminogen) -based carrier, MTPABP-Guided-Intracellular-Carrier (MAGIC), that achieves high efficiency in delivering native proteins into the cytosol. MAGIC is capable of binding both negatively and positively charged proteins and mediates their intracellular transport via clathrin-mediated uptake with endosomal escape.Results: Notably, the β-Gal and HRP proteins delivered by MAGIC maintain their biological activity after delivery, and the cytotoxicity of MAGIC is approximately 40% lower than that of Lipo8000 and TranEX. MAGIC efficiently delivered trypsin, RNase A, and saporin into the cytosol of mammalian cell lines while preserving their enzymatic or functional activity.Conclusion: This rationally designed AIEgen-based platform provides a versatile and efficient solution for cytosolic protein delivery. The MAGIC delivery system holds significant promise for advancing the study of cellular physiology and enabling precise therapeutic interventions. Keywords: AIEgen carrier, cytosolic delivery, protein transport, therapeutic proteins