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ＤＮＡナノテクノロジーを基盤とした生体分子反応の直接１分子観察と応用

基于DNA纳米技术的生物分子反应直接单分子观察及应用

基本信息

批准号：
13J01399
负责人：
楊泱泱
金额：
$ 1.03万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2013
资助国家：
日本
起止时间：
2013-04-01 至 2015-03-31
项目状态：
已结题

项目摘要

(1) Remotely controllable DNA nanocapsule for the delivery of protein by photo regulation.A 70 nm × 10 nm × 10 nm 3D DNA capsule was designed and constructed, which containing a small nanospace inside employed for carrying target molecules. The nanostructure has been successfully assembled and confirmed by AFM. Here caspase-3 was chosen as target for delivery, which can induce the apoptosis of cancer cells. The expression of the protein has been successfully carried out and the activity also confirmed.The caspase-3 labeled nanocapsule will be further introduced into the cell system. The photoinduced cleavage of caged nanocapsule has been examined by AFM before and after the labeling.(2) Direct visualization of rotational motions on a single DNA origamiA novel nanomechanical device based on DNA origami was developed by empoying DNA photoswitches. A rectangular DNA origami assembled with two double-crossover molecules connected by a rigid linker: a short nucleic acid strand modified with oligophenylene-ethylene, in which one of the DX molecules was located on the surface of DNA tile as a pointer. The nanostructures were constructed and confirmed by AFM. Two pairs of photoresponsive oligonucleotides containing azobenzene molecules were introduced into DX pointer and DNA tile scaffold separately. By switching the photoirradiation with different wavelengths, the DX pointer can be driven to rotate on the DNA tile in a pointer-on-a clock fashion. Moreover, reversible rotational movement of DX pointer on single DNA tile was directly visualized using high-speed AFM.

(1)通过光调控实现蛋白质递送的远程可控DNA纳米胶囊。设计并构建了一个70 nm × 10 nm × 10 nm的三维DNA胶囊，胶囊内含有一个小的纳米空间，用于携带目标分子。成功组装了纳米结构，并通过原子力显微镜进行了验证。本研究选择caspase-3作为递送靶点，caspase-3可以诱导癌细胞凋亡。该蛋白的表达已成功进行，活性也得到了证实。caspase-3标记的纳米胶囊将进一步引入细胞系统。用原子力显微镜观察了贴标前后笼型纳米胶囊的光致裂现象。(2)单DNA折纸旋转运动的直接可视化利用DNA光开关，研制了一种基于DNA折纸的新型纳米机械装置。一个长方形的DNA折纸，由两个双交叉分子通过刚性连接物连接而成：一条短的核酸链被聚苯乙烯修饰，其中一个DX分子位于DNA瓦片表面作为指针。通过原子力显微镜对纳米结构进行了构建和验证。将两对含偶氮苯分子的光响应寡核苷酸分别引入DX指针和DNA瓦支架中。通过切换不同波长的光照射，可以驱动DX指针以指针对时钟的方式在DNA瓦片上旋转。利用高速原子力显微镜直接观察DX指针在单个DNA瓦片上的可逆旋转运动。