Nanoenhancer for improving naked DNA electrotransfection In vivo.
Nanoenhancer for improving naked DNA electrotransfection In vivo.
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DOI：
10.3389/fbioe.2023.1181795
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发表时间：
2023
影响因子：
5.7
通讯作者：
Yuan, Fan
中科院分区：
文献类型：
--
作者：
Wang, Yifei;Wang, Chunxi;Sylvers, Justin;Segura, Tatiana;Yuan, Fan
引言：电转染（ET）是一种广泛用于递送裸核酸的非病毒方法。其效率可以通过用各种小分子增强剂处理细胞而在体外增加。然而，这些增强剂通常无法改善体内ET，这可能是由于局部注射后组织中的快速清除，减少了它们的细胞吸收。为此，我们建议开发一种新型的ET增强剂，我们称之为nanoenhancer，它在组织中扩散缓慢，不易被血液和淋巴微血管吸收。 方法：以海藻酸钠（Alg）和壳聚糖（Chi）为原料，分别与聚乙烯亚胺（PEI）或聚乙烯亚胺（PEI）复合，制备两种纳米增强剂。在编码报告基因的质粒DNA的ET之前，它们被用于体外处理细胞或体内处理小鼠后腿肌肉。在ET后24小时，量化ET的效率，并与未处理的对照组进行比较。用荧光显微镜技术测量纳米增强剂处理后溶酶体大小和酸度的变化。 结果和讨论：我们观察到用纳米增强剂预处理细胞可以提高C2C12和HCT 116细胞的ET效率和细胞活力，并且纳米增强剂预处理对体内ET效率具有相似的影响。增强的机制与纳米增强剂处理引发的溶酶体功能的瞬时失活有关。纳米增强剂的概念将导致开发新的增强剂，可用于提高体内ET效率，突出其在临床应用中的潜力。
Introduction: Electrotransfection (ET) is a non-viral approach widely used for delivery of naked nucleic acids. Its efficiency can be increased in vitro by treatment of cells with various small molecule enhancers. However, these enhancers often fail to improve ET in vivo, presumably due to rapid clearance in tissues after local injection, reducing their cellular uptake. To this end, we propose to develop a new type of ET enhancers, which we term nanoenhancer, that diffuse slowly in tissues and are poorly absorbed by blood and lymph microvessels. Methods: Two nanoenhancers were synthesized with alginate (Alg) and chitosan (Chi) with or without poly (ethylene imine) (PEI). They were used to treat cells in vitro or mouse muscle in the hind leg in vivo prior to ET of plasmid DNA coding reporter genes. At 24 hours post ET, the efficiency of ET was quantified, and compared with that in the untreated controls. Changes in lysosomal size and acidity post nanoenhancer treatment were measured with fluorescence microscopy techniques. Results and discussion: We observed that the pretreatment of cells with the nanoenhancers could enhance the ET efficiency and cell viability in both C2C12 and HCT116 cells in vitro, and the nanoenhancer pretreatment had similar effects on the ET efficiency in vivo. Mechanisms of the enhancement were related to transient inactivation of lysosomal functions triggered by the nanoenhancer treatment. The concept of nanoenhancer will lead to development of new enhancers that can be used to improve ET efficiency in vivo, highlighting its potential in clinical applications.
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影响因子：
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作者：
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