Xiao Chenglu, Wang Fang, Hou Junjie, Zhu Xiaojun, Luo Ying, Xiong Jing-Wei
Beijing Key Laboratory of Cardiometabolic Molecular Medicine, State Key Laboratory of Natural and Biomimetic Drugs, Institute of Molecular Medicine, Peking University.
Department of Biomedical Engineering, College of Engineering, Peking University.
J Vis Exp. 2018 Jul 29(137):58054. doi: 10.3791/58054.
Mammals have a very limited capacity to regenerate the heart after myocardial infarction. On the other hand, the adult zebrafish regenerates its heart after apex resection or cryoinjury, making it an important model organism for heart regeneration study. However, the lack of loss-of-function methods for adult organs has restricted insights into the mechanisms underlying heart regeneration. RNA interference via different delivery systems is a powerful tool for silencing genes in mammalian cells and model organisms. We have previously reported that siRNA-encapsulated nanoparticles successfully enter cells and result in a remarkable gene-specific knockdown in the regenerating adult zebrafish heart. Here, we present a simple, rapid, and efficient protocol for the dendrimer-mediated siRNA delivery and gene-silencing in the regenerating adult zebrafish heart. This method provides an alternative approach for determining gene functions in adult organs in zebrafish and can be extended to other model organisms as well.
哺乳动物在心肌梗死后心脏再生的能力非常有限。另一方面，成年斑马鱼在进行心尖切除或冷冻损伤后能够再生其心脏，这使其成为心脏再生研究的重要模式生物。然而，缺乏针对成年器官的功能丧失方法限制了我们对心脏再生潜在机制的深入了解。通过不同递送系统进行的RNA干扰是在哺乳动物细胞和模式生物中沉默基因的有力工具。我们之前曾报道，包裹小干扰RNA（siRNA）的纳米颗粒能够成功进入细胞，并在成年斑马鱼再生心脏中导致显著的基因特异性敲低。在此，我们展示了一种简单、快速且高效的方案，用于在成年斑马鱼再生心脏中进行树枝状大分子介导的siRNA递送和基因沉默。该方法为确定斑马鱼成年器官中的基因功能提供了一种替代方法，并且也可以扩展到其他模式生物。
