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

有了这个奖项，数学和物理科学理事会化学部门的生命过程化学项目，生物科学理事会综合有机体系统部门的发展系统集群，以及工程理事会的cbet -纳米生物传感项目将资助斯坦福大学的James Chen博士开发基于镧系元素的系统，用于体内rna和蛋白质的成像。结合镧系螯合物并增强其光致发光的RNA适体和多肽受体将通过反复的分子进化、选择和扩增来开发。受体/螯合物对随后将被用于检测活斑马鱼胚胎中亚纳摩尔水平的mrna和蛋白质，利用镧系化合物长发光寿命的优势。结合使用转录激活因子样核酸酶的基因组编辑，这些技术有可能同时观察组织模式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，这项研究将有助于解码产生复杂生物体的动态遗传程序。虽然目前胚胎发育可以用单细胞分辨率的延时显微镜观察到，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rna和蛋白质如何促进不同组织和器官形成的理解是有限的。本研究开发的新技术可以通过实时检测活斑马鱼胚胎和其他生物体中的rna和蛋白质来克服这一障碍。该方法是基于使用不同相关学科的方法，特别是合成化学、分子成像和发育生物学。培养一批科学多语种的研究生和博士后骨干。该项目还将促进发展中国家科学界的跨学科研究，方法是在巴西圣保罗大学开设化学生物学短期课程，并让有才华的巴西研究生到斯坦福大学进行研究。

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.