Vaults as Nonviral Nucleic Acid Delivery Vehicles

Vault 作为非病毒核酸递送载体

• 批准号: 7365241
• 负责人: LEONARD H ROME
• 金额: $ 32.51万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7365241
• 关键词: Baculoviruses; Binding Sites; Biological; Cells; Chemicals; Complementary DNA; Cultured Cells; DNA; Electroporation; Encapsulated; Engineering; Genes; Immune response; Insecta; Lipids; Molecular; Monitor; N-terminal; Nuclear; Nucleic Acid Binding; Nucleic Acids; Peptides; Production; Proteins; Reagent; Ribosomes; Signal Transduction; Structural Protein; Structure; System; Techniques; Technology; Tertiary Protein Structure; Testing; Time; Translations; Untranslated RNA; Viral; Virus; base; capsule; cell type; design; gene therapy; interest; nanoscale; neoplastic cell; novel strategies; particle; receptor mediated endocytosis; size; targeted delivery; uptake; vector

DESCRIPTION (provided by applicant): Gene therapy requires affordable, scaleable technology for the specific delivery of a gene of interest to the nuclear compartment of a target cell without triggering an immune response. Although viral and nonviral vectors have been designed to deliver DNA to cells, interest in nonviral vectors has grown from the promise that they can overcome numerous limitations inherent in manipulating viruses including their complexity, difficulty to adapt to large-scale production and ease of triggering an immune response. The nonviral approach described in this proposal, the engineering of a naturally-occurring nano scale structure, is unique and largely unprecedented. However, given the significant challenges that must be overcome before successful gene therapy can be achieved, a bold new approach is warranted. The vault particle is a highly conserved, abundant and ubiquitous nanocapsule with a large internal capacity to encapsulate and protect labile biological compounds such as nucleic acids. Although the 13 MDa mass of the particle makes it nearly four times the size of the ribosome, the particle is composed of multiple copies of just three proteins and one or more small untranslated RNAs. One of these proteins, MVP, is the major structural protein of the particle. When expressed in insect cells, 96 copies of MVP can assemble into the basic vault capsule. Molecular manipulation of the MVP cDNA allows packaging of specific peptides and proteins into the vault lumen. This proposal outlines a strategy to manipulate the vault nanocapsule so that it can be used to specifically package nucleic acids. In addition, an approach is delineated to modify the particle with cell targeting signals that will allow the specific targeting of these structures to normal or neoplastic cells.

描述（由申请人提供）：基因治疗需要负担得起的可缩放技术，以使目标细胞的核腔室的特定递送，而无需触发免疫反应。尽管已经设计了病毒和非病毒载体将DNA输送到细胞中，但对非病毒载体的兴趣已经源于承诺，即他们可以克服操纵病毒固有的许多局限性，包括其复杂性，难以适应大规模生产和易于触发免疫反应的易害。在本提案中描述的非病毒方法是自然存在的纳米尺度结构的工程，是独特的，并且在很大程度上是前所未有的。但是，鉴于在成功实现基因疗法之前必须克服的重大挑战，因此有必要采取一种大胆的新方法。金库颗粒是一种高度保守的，丰富的和普遍存在的纳米胶囊，内部能力较大，可以封装和保护不稳定的生物化合物，例如核酸。尽管粒子的13个MDA质量使其成为核糖体大小的几乎四倍，但该粒子仅由仅三个蛋白质的多个副本和一个或多个小的未翻译RNA组成。这些蛋白质之一MVP是粒子的主要结构蛋白。当在昆虫细胞中表达时，96份MVP副本可以组装到基本的金库胶囊中。 MVP cDNA的分子操作使特定肽和蛋白质包装到保管腔中。该建议概述了操纵金库纳米胶囊的策略，以便它可以用来专门包装核酸。另外，描述了一种方法，以通过细胞靶向信号来修改粒子，该信号将允许这些结构对正常或肿瘤细胞的特定靶向。

项目成果

Vaults are dynamically unconstrained cytoplasmic nanoparticles capable of half vault exchange.

穹窿是动态不受约束的细胞质纳米颗粒，能够进行半穹窿交换。

• DOI: 10.1021/nn102051r
• 发表时间: 2010
• 期刊: ACS nano
• 影响因子: 17.1
• 作者: Yang,Jian;Kickhoefer,ValerieA;Ng,BennyC;Gopal,Ajaykumar;Bentolila,LaurentA;John,Scott;Tolbert,SarahH;Rome,LeonardH
• 通讯作者: Rome,LeonardH

Vault nanoparticles engineered with the protein transduction domain, TAT48, enhances cellular uptake.

采用蛋白质转导结构域 TAT48 设计的 Vault 纳米粒子可增强细胞摄取。

• DOI: 10.1039/c2ib20119d
• 发表时间: 2013
• 期刊: Integrative biology : quantitative biosciences from nano to macro
• 作者: Yang,Jian;Srinivasan,Aswin;Sun,Yang;Mrazek,Jan;Shu,Zhanyong;Kickhoefer,ValerieA;Rome,LeonardH
• 通讯作者: Rome,LeonardH