Engineering Bioimaging Probes Based on Triggered Molecular Geometry

描述（由申请人提供） 翻译后摘要：显微镜是研究生物学的核心工具。显微技术的核心是成像探针，它将不可见的生物信息转换为可成像的信号。目前的成像探针主要是通过操纵它们的光学、化学和物理性质来设计的。但现代显微镜在分辨形状方面的能力不断增强，而纳米技术在精确设计形状方面的能力正在为基于“工程几何学”的新型探针创造一个惊人的机会。我们建议工程成像探针的基础上“触发分子几何”：在检测到分子信号，探针产生一个规定的，可成像的分子形状。我们的探针由亚稳态核酸单体组成。只有在检测到目标生物分子时，它们才能自主自组装成具有规定几何形状的可编程3D结构。形状可以直接成像;或者，它可以作为其他成像实体的空间组织者或放大方案。为了实现这一目标，我们将基于我们最近在工程化核酸自组装途径方面的成功来开发合成分子自组装的新范例，其中合成分子结构遵循明确设计的动力学途径等温生长。这种新的范例与目前基于热退火的范例在根本上不同，并且在概念上比基于热退火的范例更强大，并且独特地适合于实现用于生物医学应用的触发分子几何结构。为了证明其能力，我们将构建用于三种使能生物成像应用的探针：（1）用于同时成像许多不同mRNA种类的触发荧光几何条形码，（2）将使目前的荧光蛋白技术更加灵活、表达力更强且更易于使用的蛋白质组织者，以及（3）触发几何条形码作为电子冷冻断层扫描的长期受欢迎的视觉标记。如果成功，拟议的工作将允许几何学加入光学，化学和物理学，作为构建成像探针的基本，工程性质，具有深远的科学和医学意义。 公共卫生相关性：我们建议基于触发分子几何结构设计生物成像探针。探针在检测到目标生物分子信号时产生规定的可成像3D形状。如果成功，拟议的工作将允许几何学加入光学，化学和物理学，作为构建成像探针的基本，工程性质，具有深刻的科学和医学意义。

DESCRIPTION (Provided by the applicant) Abstract: Microscopy is a central tool for studying biology. At the heart of microscopic technology is the imaging probe, which transduces invisible biological information to an imageable signal. Present imaging probes are mostly engineered by manipulating their optical, chemical, and physical properties. But modern microscopy's ever in- creasing power to resolve shape, and nanotechnology's power to precisely engineer shape are creating a striking opportunity for a new kind of probes based on "engineered geometry". We propose to engineer imaging probes based on "triggered molecular geometry": upon detecting a molecular signal, the probe produces a prescribed, imageable molecular shape. Our probe consists of metastable nucleic acid monomers. Only upon detecting a target biomolecule, they autonomously self-assemble into a programmable 3D structure with prescribed geome- try. The shape can be imaged directly; alternatively, it can serve as a spatial organizer or amplification scheme for other imaging entities. To achieve this goal, we will build on our recent success in engineering nucleic acid self- assembly pathways to develop a new paradigm in synthetic molecular self-assembly, where a synthetic molecular structure grows isothermally, following explicitly designed kinetic pathways. This new paradigm is fundamen- tally different from and conceptually more powerful than the present thermal annealing based paradigm, and is uniquely suitable for implementing triggered-molecular-geometry for biomedical applications. To demonstrate its power, we will build probes for three enabling bioimaging applications: (1) a triggered fluorescent geometric bar- code for the simultaneous imaging of many distinct mRNA species, (2) a protein organizer that will make present fluorescent protein technology much more flexible, expressive, and easier to use, and (3) a triggered geometric barcode as a long sought-after visual marker for electron cryotomography. If successful, the proposed work will allow geometry to join optics, chemistry, and physics as a fundamental, engineerable property for constructing imaging probes, with profound scientific and medical implications. Public Health Relevance: We propose to engineer bioimaging probes based on triggered molecular geometry. The probe produces a prescribed, imageable 3D shape upon detecting a target biomolecular signal. If successful, the proposed work will allow geometry to join optics, chemistry, and physics as a fundamental, engineerable property for constructing imaging probes, with profound scientic and medical implications.