Self-assembling Nanostructures from An Expanded Genetic Information System (AEGIS)

在有机化学中，DNA和RNA在广泛的分子结构中显示角色基自组装，这在聚合物中几乎是独一无二的。这种自组装是生物技术和遗传学的基础，也是本提案的重点。本纳实验室的工作表明，沃森-克里克碱基对可以支持12个核碱基连接在6个不同的碱基对中，每个碱基对由不同的氢键模式连接，从而提供一个扩展的遗传信息系统（AEGIS），遵循扩展的沃森-克里克配对规则集。AEGIS可以通过DNA和RNA聚合酶催化的模板定向聚合来复制。Tan实验室的工作已经开发并利用了近场扫描光学显微镜（NSOM）工具，以10- 20纳米分辨率在二维表面上成像单分子，特别是具有单分子尺寸的纳米结构功能探针。利用这些技术，Tan已经对各种分子和纳米结构（巴克敏斯特富勒烷、DNA链和细胞膜）进行了成像，光子制造了各种生物化学功能化的纳米结构，分析了单个酶分子的动力学，并探测了单个细胞内代谢物的释放。这两个研究小组位于相邻的建筑中，因此他们的技术非常适合进行一个合作项目，重点关注通过生物自组装纳米结构合成/制造的实际方面。将探索使用生物催化剂（特别是耐热DNA聚合酶）合成纳米结构成分的程序，并着眼于纳米结构成分的原位生成。最大的纳米网络的细胞将装载可被NSOM写入/读取的光反应基团。我们希望这将创建一个与实际应用明显相关的小型可读/可写内存元件。

DNA and RNA are virtually unique among polymers in organic chemistry to display role-base self-assembly over a broad range of molecular structure. This self-assembly is the basis of biotechnology and genetics, and is the focus of this proposal. Work in the Benner laboratories has shown that the Watson-Crick base pair can support 12 nucleobases joined in six distinct base pairs, each joined by different patterns of hydrogen bonding, to give An Expanded Genetic Information System (AEGIS) following an expanded set of Watson- Crick pairing rules. AEGIS can be copied via template-directed polymerization catalyzed by DNA and RNA polymerases. Work in the Tan laboratories has developed and exploited near-field scanning optical microscopy (NSOM) tools to image single molecules with 10- 20 nm resolution on two-dimensional surfaces, in particular, nanostructure functional probes having the dimensions of single molecules. Using these technologies, Tan has imaged a variety of molecules and nanostructures (buckminsterfulleranes, DNA chains, and cell membranes), photonanofabricated a variety of biochemically functionalized nanostructures, analyzed the kinetics of single enzyme molecules, and probed the release of metabolites inside single cells. The two research groups, housed in adjacent buildings, therefore have technologies ideally matched to undertake a collaborative project focusing on the practical aspects of synthesis/fabrication through bio-self assembly of nanostructures. Procedures for synthesizing the components of the nanostructures using biocatalysts (in particular, thermostable DNA polymerases) will be explored, with an eye towards in situ generation of the components of nanostructures. The cells of the largest nano-nets will be loaded with photoreactive groups that can be written/read by NSOM. We hope that this will create a small readable/writable memory element with obvious relevance to practical applications.