Controlling DNA Condensation In Vitro

体外控制 DNA 缩合

• 批准号: 6727574
• 负责人: NICHOLAS V. HUD
• 金额: $ 22.7万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6727574
• 关键词: DNA; chemical condensation; gel electrophoresis; ionic strengths; nucleic acid structure; oligonucleotides; protamines; spermidine; transmission electron microscopy; triple helix

DESCRIPTION (provided by applicant): The use of DNA as a therapeutic agent holds great promise for the treatment of genetic disorders and acquired diseases. To this end, nonviral (or artificial) gene delivery systems are attractive because they can circumvent some problems and risks associated with viral gene delivery. However, the efficiency of gene delivery in vivo by artificial systems is presently much lower than that achieved by viruses. It is widely appreciated that DNA condensation (or compaction) is an important component of most approaches to gene delivery. For example, packaging DNA into particles with dimensions smaller than 50 nm would greatly facilitate its diffusion through the intercellular matrix of tissues. Multivalent cations (e.g. spermidine) can cause DNA in solution to collapse into tightly packed toroidal and rod-like particles with overall dimensions of 100-200 nm. The ability to produce DNA particles with smaller and more uniform dimensions could be very useful in the development of nonviral approaches to gene delivery. The goal of this research is to develop methods for controlling the size and shape of particles into which DNA is condensed. Dr. Hud proposes the use of localized static bends, loops and ovals to nucleate condensation along otherwise linear DNA molecules upon their condensation by multivalent cations. It is expected that static loops will promote the formation of toroidal DNA particles with dimensions governed by the diameter of the loop. Alternatively, static oval structures may favor DNA compaction into rod-like particles where rod length is influenced by the length of the oval. This research should demonstrate the extent to which DNA static structures can be used to control DNA condensation. Dr. Hud's results will have direct implications on the feasibility of using static DNA structures in the development of nonviral gene delivery systems.

描述（由申请人提供）：DNA作为治疗剂的用途 对治疗遗传性疾病和获得性疾病有很大的希望。 疾病为此，非病毒（或人工）基因递送系统被广泛应用。 吸引力，因为他们可以规避一些问题和风险， 病毒基因递送。然而，体内基因递送的效率， 人工系统目前比病毒所达到的要低得多。是 广泛认识到，DNA凝聚（或压缩）是一个重要的 是大多数基因传递方法的组成部分。例如，将DNA包装成 尺寸小于50 nm的颗粒将极大地促进其 通过组织的细胞间基质扩散。 多价阳离子（例如亚精胺）可导致溶液中的DNA塌陷 形成紧密堆积的环形和棒状颗粒， 100-200 nm。产生更小更均匀的DNA颗粒的能力 尺寸可能是非常有用的非病毒方法的发展， 基因传递这项研究的目的是开发控制方法， DNA凝聚成的颗粒的大小和形状。赫德博士提议 使用局部静态弯曲、环和椭圆形来成核冷凝 沿着另外的线性DNA分子在它们被多价 阳离子。预计静态循环将促进 环形DNA颗粒，其尺寸由环的直径决定。 或者，静态的椭圆形结构可能有利于DNA压缩成棒状结构。 其中杆长度受椭圆长度影响的颗粒。这 研究应该证明DNA静态结构可以在多大程度上被 用来控制DNA浓缩哈德医生的结果将直接 对使用静态DNA结构的可行性的影响 非病毒基因传递系统的发展。