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首页> 外文期刊>Acta biomaterialia >Multilayer mediated forward and patterned siRNA transfection using linear-PEI at extended N/P ratios.
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Multilayer mediated forward and patterned siRNA transfection using linear-PEI at extended N/P ratios.

机译：使用线性PEI以扩展的N / P比进行多层介导的正向和模式siRNA转染。

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Gene delivery from a substrate depends, in part, on the vector-nucleic acid complex that is bound to the surface and the cell adhesive properties of the surface. Here, we present a method to deliver patterns of small interfering RNA (siRNA) that capitalize on a forward transfection method (transfection by introducing siRNA transfection reagent complexes onto plated cells); herein denoted as multilayer mediated forward transfection (MFT). This method separates the substrate-mediated delivery from the cell adhesive properties of the surface. pH responsive layer-by-layer (LbL) assembled multilayers were used as the delivery platform and microcontact printing technique (microCP) was used to pattern nanoparticles of transfection reagent-siRNA complexes onto degradable multilayers. Efficient MFT depend on optimal formulation of the nanoparticles. 25 kDa linear polyethylenimine (LPEI) was optimized as the siRNA transfection reagent for normal forward transfection (NFT) of the nanoparticles. A broad range of LPEI-siRNA nitrogen/phosphate (N/P) ratios (ranging from 5 to 90) was evaluated for the relative amounts of siRNA incorporated into the nanoparticles, nanoparticle size and NFT efficiencies. All the siRNA was incorporated into the nanoparticles at N/P ratio near 90. Increasing the amount of siRNA incorporated into the nanoparticles, with increasing N/P ratio correlated with a linear blue shift in the ultraviolet/visible (UV/vis) absorbance spectrum of the LPEI-siRNA nanoparticles. NFT efficiency greater than 80% was achieved with minimal cytotoxicity at N/P ratio of 30 and siRNA concentration of 200 nM. Similarly, MFT efficiency 60% was achieved for LPEI-siRNA nanoparticles at N/P ratios greater than 30.
机译：从底物的基因递送部分取决于结合至表面的载体-核酸复合物以及该表面的细胞粘附特性。在这里，我们提出一种利用正向转染方法（通过将siRNA转染试剂复合物引入平板细胞进行转染）来传递小干扰RNA（siRNA）模式的方法;本文中称为多层介导的正向转染（MFT）。该方法将底物介导的递送与表面的细胞粘附特性分开。 pH响应层（LbL）组装的多层用作输送平台，微接触印刷技术（microCP）用于将转染试剂-siRNA复合物的纳米颗粒图案化到可降解的多层上。高效的MFT取决于纳米颗粒的最佳配方。优化25 kDa线性聚乙烯亚胺（LPEI）作为用于纳米粒子正常正向转染（NFT）的siRNA转染试剂。对于掺入纳米颗粒中的siRNA的相对量，纳米颗粒大小和NFT效率，评估了范围很广的LPEI-siRNA氮/磷酸盐（N / P）比（从5到90）。所有siRNA都以接近90的N / P比掺入纳米颗粒。增加掺入纳米颗粒的siRNA量，随着N / P比例的增加与紫外/可见（UV / vis）吸收光谱中的线性蓝移相关LPEI-siRNA纳米颗粒。在30的N / P比和200 nM的siRNA浓度下，具有最小的细胞毒性，可实现大于80％的NFT效率。同样，N / P大于30的LPEI-siRNA纳米颗粒的MFT效率达到60％。

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