Controlling DNA Condensation In Vitro

体外控制 DNA 缩合

• 批准号: 6880092
• 负责人: NICHOLAS V. HUD
• 金额: $ 22.66万
• 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 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6880092
• 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(申请人提供)：将DNA用作治疗剂 为遗传疾病的治疗带来了巨大的希望，并获得了 疾病。为此，非病毒(或人工)基因传递系统是 具有吸引力，因为它们可以规避与以下方面相关的一些问题和风险 病毒基因传递。然而，体内基因传递的效率通过 人工系统目前比病毒实现的要低得多。它是 人们普遍认识到DNA凝聚(或压缩)是一种重要的 这是大多数基因传递方法的组成部分。例如，将DNA包装到 尺寸小于50纳米的粒子将极大地促进其 通过组织的细胞间基质扩散。 多价阳离子(如亚精胺)可导致溶液中的DNA崩溃 形成紧密堆积的环状和棒状颗粒，总尺寸为 100-200 nm。生产更小、更均匀的DNA颗粒的能力 维度在开发非病毒式方法方面可能非常有用 基因传递。这项研究的目标是开发控制的方法 DNA凝聚成的颗粒的大小和形状。胡德博士建议 使用局部静态弯曲、环路和椭圆形来使凝结成核 沿其他线形DNA分子以多价凝聚 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压实成棒状 其中杆长受椭圆形长度影响的粒子。这 研究应该证明DNA静态结构可以在多大程度上 用来控制DNA缩合。哈德博士的结果将直接 关于使用静态DNA结构的可行性的暗示 开发非病毒基因传递系统。

项目成果

Integration host factor (IHF) dictates the structure of polyamine-DNA condensates: implications for the role of IHF in the compaction of bacterial chromatin.

• DOI: 10.1021/bi8019965
• 发表时间: 2009-01
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Tumpa Sarkar;A. Petrov;Jason R Vitko;C. Santai;S. Harvey;I. Mukerji;N. Hud

Complete disproportionation of duplex poly(dT)*poly(dA) into triplex poly(dT)*poly(dA)*poly(dT) and poly(dA) by coralyne.

• DOI: 10.1093/nar/30.4.983
• 发表时间: 2002-02
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Matjaž Polak;N. Hud

Formation of native-like mammalian sperm cell chromatin with folded bull protamine.

与折叠的公牛鱼精蛋白形成类似天然的哺乳动物精子细胞染色质。

• DOI: 10.1074/jbc.m312777200
• 发表时间: 2004
• 期刊: The Journal of biological chemistry
• 作者: Vilfan,IgorD;Conwell,ChristineC;Hud,NicholasV
• 通讯作者: Hud,NicholasV

Bacterial protein HU dictates the morphology of DNA condensates produced by crowding agents and polyamines.

• DOI: 10.1093/nar/gkl1093
• 发表时间: 2007
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Sarkar T;Vitoc I;Mukerji I;Hud NV
• 通讯作者: Hud NV