Department of Bioengineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan.
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Chiyoda, Tokyo 101-0062, Japan.
J Mater Chem B. 2021 Jun 3;9(21):4298-4302. doi: 10.1039/d1tb00645b.
Endosomal escape is crucial for the delivery of nucleic acids. However, the understanding of the underlying mechanisms is still deficient. In this work, we explored the effects of lipid- and polymer-based transfection reagents on the permeability of cellular membranes through an innovative method combining a proton-sensing transistor and a cytosolic LDH leakage assay, which allows us to distinguish between modes of molecule permeation that may occur during endosomal escape. By testing the commercial reagents lipofectin and in vivo JetPEI under physiological and endosomal pH conditions, we found that both lipid- and polymer-based transfection reagents have pH-dependent pore-forming activity, with the former creating smaller pores than the latter. This versatile approach of assessing carrier-membrane interactions is expected to contribute to the development of next-generation nucleic acid delivery systems.
内涵体逃逸对于核酸的递送至关重要。然而，对于其潜在机制的理解仍然不足。在这项工作中，我们通过一种创新的方法结合质子敏感晶体管和细胞质 LDH 渗漏测定，探索了脂质体和聚合物类转染试剂对细胞膜通透性的影响，该方法可以区分可能发生在内涵体逃逸过程中的分子渗透模式。通过在生理和内涵体 pH 条件下测试商业试剂 lipofectin 和 in vivo JetPEI，我们发现脂质体和聚合物类转染试剂都具有 pH 依赖性的孔形成活性，前者形成的孔比后者小。这种评估载体-膜相互作用的多功能方法有望为下一代核酸递送系统的发展做出贡献。
