NER: Oligonucleotide-Directed Alignment of Cytoskeletal Filaments for Nanoscale Assembly

NER：用于纳米级组装的细胞骨架丝的寡核苷酸定向排列

• 批准号: 0209687
• 负责人: William Hancock
• 金额: $ 9.5万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-15 至 2004-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0209687&HistoricalAwards=false
• 关键词: NER; Oligonucleotide; Directed; Alignment; Cytoskeletal

NER: Oligonucleotide-directed alignment of cytoskeletal filaments for nanoscale assemblyThere is currently great interest in nanoscience and nanotechnology for manipulating material at submicron scales. In cells, however, this is a routine task -- the transport of intracellular cargo is carried out by motor proteins that move along cytoskeletal filaments called microtubules. Directional movement is achieved because microtubules have a structural polarity and motors move unidirectionally along them, with different motors moving in different directions. We seek to: (i) reconstruct the intracellular transport system in vitro using purified kinesin motor proteins and microtubules, and (ii) use it to transport defined cargo (biological or synthetic nanoscale objects) to specific sites on a two-dimensional substrate. To lay down microtubule tracks with desired orientations, we will take advantage of the specificity and reversibility of DNA hybridization as a "molecular glue". Single-stranded DNA oligonucleotides will be attached to defined sites on glass surfaces and complementary oligonucleotides will be covalently attached to microtubules. By making microtubules whose ends are functionalized with different DNA sequences and spatially patterning their sequence complements, a variety of microtubule geometries will be investigated. These tracks will be used to direct the motion of cargo-laden kinesin motors as a step towards building systems for biomolecular separations or directed assembly applications.

NER：纳米级装配的细胞骨架细丝的寡核苷酸指导对准目前对在亚微米尺度上操纵材料的纳米科学和纳米技术对纳米科学和纳米技术非常感兴趣。 然而，在细胞中，这是一个常规任务 - 细胞内货物的运输是由沿着称为微管的细胞骨架丝移动的运动蛋白进行的。 由于微管具有结构性极性，并且电动机沿它们单向移动，因此可以实现方向运动，并且电动机朝不同的方向移动。 我们寻求：（i）使用纯化的驱动蛋白运动蛋白和微管在体外重建细胞内转运系统，并（ii）使用它来将定义的货物（生物或合成纳米级物体）运输到二维底物上的特定位点。 为了将微管轨道放下所需的方向，我们将利用DNA杂交作为“分子胶”的特异性和可逆性。 单链DNA寡核苷酸将连接到玻璃表面上的定义位点，互补的寡核苷酸将共价附加到微管上。 通过制作其末端通过不同的DNA序列官能化并在空间上模仿其序列互补的微管，将研究各种微管几何形状。 这些轨道将用于指导载有货物的驱动蛋白电动机的运动，以此作为建立生物分子分离或定向装配应用的系统的一步。