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<p>RNA has become crucial in gene therapy by enabling direct and temporary therapeutic protein production, eliminating the risk of genome integration, and avoiding the insertional mutagenesis associated with DNA-based approaches. In this study, we evaluated polyethylenimine (PEI) derivatives as mRNA carriers for delivery. Modifying a kanamycin core with low-molecular-weight polyethylenimine, known as KANPEI, which was previously shown to enhance plasmid DNA transfection efficiency and reduce cytotoxicity. The ability of KANPEI derivatives to complex with mRNA was evaluated using gel electrophoresis, RiboGreen assays, and field-emission scanning electron microscopy (FE-SEM), which revealed the formation of nanosized polyplexes. The physicochemical characteristics of the resulting KANPEI/mRNA polyplexes were further characterized by dynamic light scattering and zeta potential measurements. The pH-buffering capacities of KANPEI was assessed by acid–base titration. In addition, transfection efficiency and cytotoxicity were systematically evaluated in HepG2, HeLa, and NIH3T3 cell lines. Compared with PEI 25&#xa0;kDa, KANPEIs showed markedly lower cytotoxicity while maintaining comparable or superior transfection efficiency, demonstrating their potential as effective and biocompatible mRNA delivery carriers.</p> Graphical abstract <p></p>

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Synthesis and evaluation of kanamycin–polyethylenimine conjugates as efficient and biocompatible mRNA carriers

  • 期刊论文

  • 发表日期：

    2026年2月20日

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  • Seongyeon Kim,
  • Onesun Jung,
  • Viet Dongquoc,
  • Minyoung Choi,
  • Sangik Hwang,
  • Ji Heon Noh,
  • Le Thi Thuy,
  • Joon Sig Choi

摘要

RNA has become crucial in gene therapy by enabling direct and temporary therapeutic protein production, eliminating the risk of genome integration, and avoiding the insertional mutagenesis associated with DNA-based approaches. In this study, we evaluated polyethylenimine (PEI) derivatives as mRNA carriers for delivery. Modifying a kanamycin core with low-molecular-weight polyethylenimine, known as KANPEI, which was previously shown to enhance plasmid DNA transfection efficiency and reduce cytotoxicity. The ability of KANPEI derivatives to complex with mRNA was evaluated using gel electrophoresis, RiboGreen assays, and field-emission scanning electron microscopy (FE-SEM), which revealed the formation of nanosized polyplexes. The physicochemical characteristics of the resulting KANPEI/mRNA polyplexes were further characterized by dynamic light scattering and zeta potential measurements. The pH-buffering capacities of KANPEI was assessed by acid–base titration. In addition, transfection efficiency and cytotoxicity were systematically evaluated in HepG2, HeLa, and NIH3T3 cell lines. Compared with PEI 25 kDa, KANPEIs showed markedly lower cytotoxicity while maintaining comparable or superior transfection efficiency, demonstrating their potential as effective and biocompatible mRNA delivery carriers.

Graphical abstract