Department of Tumor Biology, Institute for Cancer Research , The Norwegian Radium Hospital, Montebello, Norway.
Nucleic Acid Ther. 2013 Apr;23(2):160-5. doi: 10.1089/nat.2012.0413. Epub 2013 Mar 26.
A promising method that offers both time- and site-specific delivery of macromolecules is photochemical internalization technology (PCI). Here, we have characterized various polyamidoamine (PAMAM) carriers [generation (G) 0-7], for light-directed delivery of nucleic acids in vitro by the use of PCI technology. A number of parameters for optimal delivery of nucleic acids into human cancer cells, that is, various light-doses, carrier-doses, and small interfering RNA (siRNA)/messenger RNA (mRNA) doses were investigated for either up- or down-regulation of enhanced green fluorescent protein (EGFP) gene expression. In summary, our results showed in an osteosarcoma cell line (OHS) [EGFP] model system the possibility for efficient light-directed siRNA silencing (>80% silencing) when using PAMAM G3 to G7 as carriers. Surprisingly, no EGFP mRNA up-regulation was detected either with or without PCI after EGFP mRNA/PAMAM (G0-G7) transfection in standard OHS cells. We have here identified properties for PAMAM formulations enabling light-directed siRNA delivery with the aim of developing a site-specific strategy for delivery of nucleic acids in vivo.
一种有前途的方法，提供了时间和特定部位的大分子传递是光化学内化技术（PCI）。在这里，我们已经描述了各种聚酰胺胺（PAMAM）载体[代（G）0-7]，用于通过使用 PCI 技术体外传递核酸。许多参数的最佳传递核酸进入人类癌细胞，即各种光剂量，载体剂量和小干扰 RNA（siRNA）/信使 RNA（mRNA）剂量被调查上调或下调增强型绿色荧光蛋白（EGFP）基因表达。总之，我们的结果表明在骨肉瘤细胞系（OHS）[EGFP]模型系统中，当使用 PAMAM G3 至 G7 作为载体时，siRNA 沉默的效率很高（>80%的沉默）。令人惊讶的是，在用或不用 PCI 后，在标准 OHS 细胞中转染 EGFP mRNA/PAMAM（G0-G7）后，没有检测到 EGFP mRNA 的上调。我们已经确定了 PAMAM 配方的特性，使 siRNA 的光控传递成为可能，目的是开发体内核酸传递的特异性策略。
