Exosomes are attractive as nucleic-acid carriers because of their favourable pharmacokinetic and immunological properties and their ability to penetrate physiological barriers that are impermeable to synthetic drug-delivery vehicles. However, inserting exogenous nucleic acids, especially large messenger RNAs, into cell-secreted exosomes leads to low yields. Here we report a cellular-nanoporation method for the production of large quantities of exosomes containing therapeutic mRNAs and targeting peptides. We transfected various source cells with plasmid DNAs and stimulated the cells with a focal and transient electrical stimulus that promotes the release of exosomes carrying transcribed mRNAs and targeting peptides. Compared with bulk electroporation and other exosome-production strategies, cellular nanoporation produced up to 50-fold more exosomes and a more than 103-fold increase in exosomal mRNA transcripts, even from cells with low basal levels of exosome secretion. In orthotopic phosphatase and tensin homologue (PTEN)-deficient glioma mouse models, mRNA-containing exosomes restored tumour-suppressor function, enhanced inhibition of tumour growth and increased survival. Cellular nanoporation may enable the use of exosomes as a universal nucleic-acid carrier for applications requiring transcriptional manipulation.

中文翻译：

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通过细胞纳米穿孔大规模生成功能性 mRNA 封装外泌体。

外泌体作为核酸载体具有吸引力，因为它们具有良好的药代动力学和免疫学特性，并且能够穿透合成药物递送载体不可渗透的生理屏障。然而，将外源核酸，尤其是大信使 RNA 插入细胞分泌的外泌体会导致产量低。在这里，我们报告了一种细胞纳米穿孔方法，用于生产大量含有治疗性 mRNA 和靶向肽的外泌体。我们用质粒 DNA 转染各种源细胞，并用局部和瞬时电刺激刺激细胞，促进释放携带转录 mRNA 和靶向肽的外泌体。与批量电穿孔和其他外泌体生产策略相比，细胞纳米穿孔产生了高达 50 倍的外泌体和超过 103 倍的外泌体 mRNA 转录物增加，即使是来自外泌体分泌基础水平低的细胞也是如此。在原位磷酸酶和张力蛋白同源物 (PTEN) 缺陷型神经胶质瘤小鼠模型中，含有 mRNA 的外泌体恢复了肿瘤抑制功能，增强了对肿瘤生长的抑制作用并提高了存活率。细胞纳米穿孔可以使外泌体作为通用核酸载体用于需要转录操作的应用。增强对肿瘤生长的抑制作用并增加存活率。细胞纳米穿孔可以使外泌体作为通用核酸载体用于需要转录操作的应用。增强对肿瘤生长的抑制作用并增加存活率。细胞纳米穿孔可以使外泌体作为通用核酸载体用于需要转录操作的应用。