Department of Chemical and Pharmaceutical Sciences, University of Trieste, Piazzale Europa 1, Trieste, 34127, Italy.
Nanoscale. 2013 Oct 7;5(19):9110-7. doi: 10.1039/c3nr02694a. Epub 2013 Aug 1.
In the present study, we report the design and synthesis of peptide-based-multi-walled carbon nanotubes (MWCNTs) to target mitochondria. Targeting these intracellular organelles might open the way to develop alternative systems to address diseases related to genetic mutations in mitochondrial (mt)-DNA, by delivering therapeutic oligonucleotides. The first step towards mitochondrial delivery of this type of nucleic acid was to target MWCNTs to mitochondria by covalent functionalization with a well-known endogenous mitochondrial targeting sequence (MTS). The subcellular localization of the conjugates, which were fluorescently labeled, in murine RAW 264.7 macrophages and human HeLa cells was then studied using different microscopy techniques, such as wide-field epifluorescence microscopy, confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). The localization of the MTS-MWCNT conjugates into mitochondria was further confirmed by analyzing the isolated organelles using TEM.
在本研究中,我们报告了基于肽的多壁碳纳米管 (MWCNTs) 的设计和合成,以靶向线粒体。靶向这些细胞内细胞器可能为开发替代系统提供了一种途径,通过递送电疗寡核苷酸来解决与线粒体 (mt)-DNA 基因突变相关的疾病。将这种类型的核酸递送到线粒体的第一步是通过与众所周知的内源性线粒体靶向序列 (MTS) 共价功能化,将 MWCNTs 靶向线粒体。然后使用不同的显微镜技术(如宽场荧光显微镜、共聚焦激光扫描显微镜 (CLSM) 和透射电子显微镜 (TEM))研究了荧光标记的缀合物在鼠 RAW 264.7 巨噬细胞和人 HeLa 细胞中的亚细胞定位。通过使用 TEM 分析分离的细胞器,进一步证实了 MTS-MWCNT 缀合物进入线粒体的定位。