Interrogation of retroelement-derived proteins for functional gene transfer

项目摘要和摘要 基因疗法通过将核酸送入体内，治疗大量的遗传性疾病。 这是疾病的根源。这是有利的，因为它是高度模块化的，允许许多不同的货核 根据致病原因输送的酸液。因此，理想的基因治疗递送载体应该能够 运载各种货物，以有针对性的方式交付，并容纳一系列货物大小。有很多种 用于传递核酸的技术，包括病毒系统如腺病毒、腺相关病毒(AAV)和 慢病毒，以及包括纳米颗粒在内的非病毒方法。尽管这些疗法可以取得成功，但一个关键 目前使用的载体的限制是免疫反应，这可能导致核酸货物的无效传递。 目前有必要开发有效的、非免疫原性的基因治疗载体，用于广泛的 疾病，包括神经系统疾病，尚未设计出有效的运送工具。 为此，哺乳动物的基因组包含许多类似病毒的基因，其中一些已经被宿主所选择 重要功能的细胞。其中有编码衣壳蛋白的Gag的同源物。我们假设 编码衣壳结构域的内源基因具有自组装成衣壳并调节细胞间的能力 通过结合、分泌和运送核酸货物进行的通讯。我们建议探索和重新设计 用作基因治疗载体的内源性衣壳蛋白。我们假设送货车辆是由 完全的自身蛋白将比标准载体更有效，因为它们可能是非免疫原性的。在这里，我们建议 使用体外鉴定、再工程和体外和体内验证相结合的方法来鉴定 并学习如何对它们进行重新设计。理想情况下，这些系统将是模块化的，同时具有 可编程的货物和取向，治疗一系列疾病。我们希望通过识别和重新设计这些 系统，由此产生的完全内生递送工具将有助于高效、可重新编程和 非免疫原性基因传递。 该项目的目标是成为一名独立的研究人员，该项目还将支持计算的开发 生物学技能、分子生物学专业知识以及导师和科学交流技能。这些将是 在博德研究所和麻省理工学院良好的研究环境的支持下。

Project Summary and Abstract Gene therapy enables the treatment of a large number of genetic diseases through delivery of nucleic acids striking at the root of the disease. This is advantageous because it is highly modular, allowing for a number of different cargo nucleic acids to be delivered depending on the disease cause. As such, the ideal gene therapy delivery vector would be able to carry a variety of cargo, deliver this in a targeted manner, and accommodate a range of cargo sizes. There are a number of techniques utilized to deliver nucleic acids including viral systems like adenovirus, adeno-associated virus (AAV), and lentivirus, as well as non-viral methods including nanoparticles. Although these therapies can be successful, a key limitation to currently used vectors is the immune response which can lead to ineffective delivery of nucleic acid cargo. There is currently a need to develop effective and non-immunogenic delivery vehicles for gene therapy for a wide range of diseases, including neurological disease, for which effective delivery vehicles have yet to be designed. To this end, mammalian genomes contain numerous virus-like genes, some of which have been co-opted by their host cells for important functions. Among these are homologs of gag, which encodes the capsid protein. We hypothesize that endogenous genes encoding a capsid domain have the ability to self-assemble into capsids and mediate intercellular communication by binding, secreting, and delivering nucleic acid cargos. We propose to explore and re-engineer endogenous capsid-containing proteins for use as gene therapy vectors. We hypothesize that delivery vehicles composed entirely of self proteins will be more effective than standard vectors as they could be non-immunogenic. Here we propose to use an approach combining in vitro characterization, re-engineering, and in vitro and in vivo validation to identify candidate proteins and learn how they can be re-engineered. These systems will ideally be modular, having both programmable cargo and tropism to treat a range of diseases. We hope that by identifying and re-engineering these systems, the resulting fully endogenous delivery vehicle will be useful for efficient, reprogrammable, and non-immunogenic gene delivery. With the goal of becoming an independent investigator, this project will also support development of computational biology skills, molecular biology expertise as well as mentorship and scientific communication skills. These will be supported by the excellent research environment at the Broad Institute and MIT.