INSPIRE Track 1: Lanthanide-based probes for visualizing RNAs and proteins in live organisms

凭借这一奖项，数学和物理科学局化学部的生命过程化学计划、生物科学局综合组织系统司的开发系统群和工程局的CBET纳米生物传感计划正在资助斯坦福大学的James Chen博士开发基于稀土的系统，用于活体RNA和蛋白质的成像。RNA适配子和多肽受体将通过反复的分子进化、选择和扩增来开发与稀土络合物结合并增强其光致发光的多肽受体。然后，利用稀土络合物的长发光寿命，受体/螯合物对将被用于检测活斑马鱼胚胎中亚纳米分子水平的mRNAs和蛋白质。与使用转录激活物样效应核酸酶的基因组编辑相结合，这些技术有可能同时观察组织构型和器官发生以及调节这些过程的大分子。通过这种跨学科的方法，这项研究将有助于破译导致复杂有机体产生的动态遗传程序。虽然目前可以通过时间推移显微镜观察到单细胞分辨率的胚胎发育，但促成这种显著转变的生物分子在很大程度上仍然是不可见的。因此，我们对特定的RNA和蛋白质如何对不同的组织和器官的形成做出贡献的了解是有限的。这项研究开发了新的技术，可以通过实时检测活斑马鱼胚胎和其他生物中的RNA和蛋白质来克服这一障碍。该方法基于与不同相关学科相关的方法的使用，特别是合成化学、分子成像和发育生物学。将培养一支科学掌握多种语言的研究生和博士后干部队伍。该项目还将通过巴西S圣保罗大学的化学生物学短期课程和让有才华的巴西研究生接触斯坦福大学的研究，促进发展中国家科学界的跨学科研究。

With this award, the Chemistry of Life Processes Program of the Chemistry Division in the Directorate for Mathematical and Physical Sciences, the Developmental Systems Cluster in the Division of Integrative Organismal Systems of the Directorate for Biological Sciences, and the CBET-Nano-Biosensing Program in the Directorate for Engineering are funding Dr. James Chen from Stanford University to develop lanthanide-based systems for the imaging of RNAs and proteins in vivo. RNA aptamers and polypeptide receptors that bind to lanthanide chelates and enhance their photoluminescence will be developed through iterative rounds of molecular evolution, selection, and amplification. The receptor/chelate pairs will then be used to detect subnanomolar levels of mRNAs and proteins in live zebrafish embryos, taking advantage of the long luminescence lifetimes of lanthanide complexes. In combination with genome editing using transcription activator-like effect nucleases, these technologies have the potential to enable the simultaneous observation of tissue patterning and organogenesis and the macromolecules that regulate these processes. Through this interdisciplinary approach, the study will help decode the dynamic genetic programs that give rise to complex organisms. While currently embryonic development can be observed with single-cell resolution by time-lapse microscopy, the biological molecules that contribute to this remarkable transformation remain largely invisible. As a result, our understanding of how specific RNAs and proteins contribute to the formation of distinct tissues and organs is limited. This study develops new technologies that may overcome this hurdle by enabling the real-time detection of RNAs and proteins in live zebrafish embryos and other organisms. The approach is based on use of methods pertaining to different related disciplines, specifically synthetic chemistry, molecular imaging, and developmental biology. A scientifically multilingual cadre of graduate students and postdoctoral fellows will be trained. The project will also promote interdisciplinary research in scientific communities from developing countries through a chemical biology short-course at the University of São Paulo in Brazil and by exposing talented Brazilian graduate students to research at Stanford University.