A universal discoidal nanoplatform for the intracellular delivery of PNAs
A universal discoidal nanoplatform for the intracellular delivery of PNAs
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DOI：
10.1039/c9nr03667a
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发表时间：
2019-07-14
期刊：
影响因子：
6.7
通讯作者：
Bahal, Raman
中科院分区：
文献类型：
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作者：
Rad, Armin Tahmasbi;Malik, Shipra;Bahal, Raman
肽核酸（PNAs）由于其在基因编辑和靶向策略中的显着潜力而获得了相当大的关注。然而，PNA的细胞递送在开发其更广泛的治疗应用方面仍然是一个挑战。在这里，我们研究了一种新的复合物的脂质bicelles和PNA为基础的载体的有效交付的PNA。为了验证概念，测试了靶向微小RNA（miR）210和155的PNA。对PNA：脂质比为1：100、1：1000和1：2500的含正电荷和负电荷的双胞进行了综合评价。PNA：脂质摩尔比为1：2500的带负电荷的双胞产生了直径约为30 nm且双层厚度为5 nm的均匀盘状形状，而带正电荷的双胞系统在掺入PNA后含有不规则囊泡。进行小角X射线散射（SAXS）分析以提供对疏水PNA如何与双胞相互作用的深入了解。此外，流式细胞术后的共聚焦显微镜分析证实了含有染料缀合的antimiR PNA的双胞的上级转染效率。功能分析还证实了通过双胞递送的PNA寡聚体的miR抑制。纳米盘状复合物开辟了一条递送PNA的新途径，PNA本身对于被内吞到细胞中是一个巨大的挑战。
Peptide nucleic acids (PNAs) have gained considerable attention due to their remarkable potential in gene editing and targeting-based strategies. However, cellular delivery of PNAs remains a challenge in developing their broader therapeutic applications. Here, we investigated a novel complex made of lipid bicelles and PNA-based carriers for the efficient delivery of PNAs. For proof of concept, PNAs targeting microRNA (miR) 210 and 155 were tested. Comprehensive evaluation of positive as well as negative charge-containing bicelles with PNA : lipid ratios of 1 : 100, 1 : 1000, and 1 : 2500 was performed. The negatively charged bicelles with a PNA : lipid molar ratio of 1 : 2500 yielded a discoidal shape with a uniform diameter of similar to 30 nm and a bilayer thickness of 5 nm, while the positively charged bicellar system contained irregular vesicles after the incorporation of PNA. Small-angle X-ray scattering (SAXS) analysis was performed to provide insight into how the hydrophobic PNAs interact with bicelles. Further, flow cytometry followed by confocal microscopy analyses substantiate the superior transfection efficiency of bicelles containing dye-conjugated antimiR PNAs. Functional analysis also confirmed miR inhibition by PNA oligomers delivered by bicelles. The nanodiscoidal complex opens a new pathway to deliver PNAs, which, on their own, are a great challenge to be endocytosed into cells.