Abstract A plethora of biological barriers, intended to defend tissues and cells against external influences, stand in the way of efficient nucleic acid delivery by non-viral nanoplexes. Even when nanoplexes successfully evade extracellular barriers and reach their target cell, many intracellular barriers remain to be conquered. These include overcoming the plasma membrane, evading endosomal compartmentalization, and in some cases crossing the nuclear envelope. At the same time, exocytosis, autophagy and cytoplasmic degradation of the cargo should be avoided. Currently, there is a growing appreciation that the interaction of nanoplexes with these barriers should be understood in detail in order to rationally design a second generation of non-viral nanoplexes, capable of overcoming these many hurdles. Studying intracellular biobarriers is, however, quite challenging and specialized methods are constantly being developed. In this review, we present an overview of established as well as emerging techniques and assays that are currently available to the experimentalist to study nanoplex-barrier interaction, with a focus on quantitative methods.

中文翻译：

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研究非病毒基因治疗中核酸递送的细胞内屏障的方法

摘要 大量旨在保护组织和细胞免受外部影响的生物屏障阻碍了非病毒纳米复合物的有效核酸递送。即使纳米复合物成功避开细胞外屏障并到达其靶细胞，仍有许多细胞内屏障有待克服。这些包括克服质膜，逃避内体区室化，以及在某些情况下穿过核膜。同时，应避免货物的胞吐作用、自噬和细胞质降解。目前，人们越来越意识到应该详细了解纳米复合物与这些障碍的相互作用，以便合理设计能够克服这些障碍的第二代非病毒纳米复合物。然而，研究细胞内生物屏障是 相当具有挑战性和专门的方法正在不断发展。在这篇综述中，我们概述了目前可供实验者研究纳米复合物-屏障相互作用的已建立和新兴技术和测定，重点是定量方法。