Non-Invasive and Non-Viral Sono-Optogenetics

项目摘要 在过去的十年里，光遗传学日益成为时空控制的一项重要技术 神经活动、心脏功能、肌肉细胞活动、蛋白质-蛋白质相互作用和疾病应用， 通过基因编码的光激活蛋白。然而，这仍然存在两大挑战 技术：1.将光传输到大脑或心脏等身体深处通常需要光学 纤维植入可能导致细胞和组织的损伤。2.基因表达需要病毒转导， 它受到一些限制，如宿主的免疫反应，稳定表达的蛋白质 随着时间的推移，对最大基因大小的限制以及缺乏制造的经济可扩展性。致信地址 第一个挑战，我们最近开发了一项名为声光遗传学的技术，将聚焦的超声波 (FUS)到光的非侵入性光遗传学。纳米粒子被注射到循环的血液中，以便 实现光遗传学既不需要开颅手术，也不需要颅内植入。然而，这些无机物 纳米颗粒通常很难被修饰成发射不同颜色的光来进行多重光遗传控制 并且在使用后在动物肝脏中积累后不能生物降解，造成长期的安全问题。 因此，这项提案的目标和我的研究实验室的重点是解决以下剩余挑战 通过设计有机纳米材料，包括氢键有机骨架的光遗传学 纳米粒子、DNA质粒的化学组装和阳离子聚合物递送剂。具体地说，我们是 计划1)设计可生物降解的纳米颗粒，将超声波转化为光，用于多种颜色的声纳- 光遗传学。2)通过共价化学组装改善通过核孔的质粒DNA的输送 策略和3)设计先进的阳离子聚合物以改善内体逃逸、细胞摄取和 非病毒基因传递中细胞外空间的扩散性。这项工作也将增进我们对 有机纳米颗粒在细胞中的传输和相互作用。因此，我相信建议的工程是 非常适合NIH R35最大化调查人员研究奖。

Project Abstract Over the past decade, optogenetics has increasingly become an important technology for spatiotemporal control of neural activity, cardio functions, muscle cell activity, protein-protein interaction, and disease applications, through the genetically encoded light-activated proteins. However, there are still two major challenges of this technology: 1.the delivery of light to into deep body areas such as brain or heart generally requires the optical fiber implantation which could result in damage of cells and tissue. 2. Gene expression requires viral transduction, which suffer from a number of limitations such as the host immune response, the stability expressed proteins over time, the limitations on maximum gene size and the lack of economic scalability for manufacture. To address the first challenge, we recently developed a technology named ‘sono-optogenetics’ to convert focused ultrasound (FUS) to light for non-invasive optogenetics. The nanoparticles are injected into the circulating blood so that neither craniotomy nor intracranial implantation is required for achieving optogenetics. However, these inorganic nanoparticles are generally difficult to be modified to emit different colors of light for multiplex optogenetic control and are not biodegradable after accumulating in the animal livers after use, causing long-term safety concerns. Therefore, the goal of this proposal and the focus of my research lab, is to tackle the remaining challenges for optogenetics through designing organic nanomaterials, including hydrogen-bonded organic frameworks nanoparticles, chemical assembly of DNA plasmids and cationic polymer delivery agents. Specifically, we are planning to 1) design biodegradable nanoparticles to convert ultrasound to light for multi-colored sono- optogenetics. 2) improve the delivery of plasmid DNAs through nucleopore through covalent chemical assembly strategies and 3) design advanced cationic polymers for improving endosome escape, cellular uptake and diffusivity through extracellular space in non-viral gene delivery. The work will also enhance our understanding the transport and interaction of organic nanoparticles in cells. As a result, I believe that the proposed works is well suited for the NIH R35 Maximizing Investigators’ Research Award.