Diao Jupeng, Wang Hongxia, Chang Nannan, Zhou Xiao-Hai, Zhu Xiaojun, Wang Jun, Xiong Jing-Wei
Institute of Molecular Medicine, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, and State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, China.
School of Life Sciences and Hefei National Laboratory for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei, China.
Dev Biol. 2015 Oct 15;406(2):196-202. doi: 10.1016/j.ydbio.2015.08.020. Epub 2015 Aug 29.
The remarkable regenerative capacity of the zebrafish has made it an important model organism for studying heart regeneration. However, current loss-of-function studies are limited by a lack of conditional-knockout and effective gene-knockdown methods for the adult heart. Here, we report a novel siRNA knockdown method facilitated by poly(ethylene glycol)-b-poly(D,L-lactide) (PEG-PLA) nanoparticles. The siRNA-encapsulated nanoparticles successfully entered cells and resulted in remarkable gene-specific knockdown in the adult heart. This effect was demonstrated by down-regulation of the Aldh1a2 and Dusp6 proteins after intrapleural delivery of nanoparticle-encapsulated siRNAs. Furthermore, siRNA-mediated knockdown of Aldh1a2 was sufficient to inhibit myocardial proliferation and decrease the numbers of Gata4-positive cardiomyocytes after ventricular resection. Therefore, the results of this work demonstrate that nanoparticle-facilitated siRNA delivery provides an alternative tool for loss-of-function studies of genes in the adult heart in particular and other organs in general in the adult zebrafish.
斑马鱼卓越的再生能力使其成为研究心脏再生的重要模式生物。然而，目前的功能丧失研究受到成体心脏缺乏条件性敲除和有效基因敲低方法的限制。在此，我们报道了一种由聚（乙二醇）-b-聚（D，L-丙交酯）（PEG-PLA）纳米颗粒辅助的新型siRNA敲低方法。包裹siRNA的纳米颗粒成功进入细胞，并在成体心脏中导致显著的基因特异性敲低。在胸腔内递送包裹纳米颗粒的siRNAs后，Aldh1a2和Dusp6蛋白的下调证明了这种效果。此外，siRNA介导的Aldh1a2敲低足以抑制心肌增殖，并减少心室切除术后Gata4阳性心肌细胞的数量。因此，这项工作的结果表明，纳米颗粒辅助的siRNA递送为成年斑马鱼成体心脏以及其他器官中基因的功能丧失研究提供了一种替代工具。
