MFB: Next-generation Proximity Labeling Technologies to Map Subcellular Transcriptomes and RNA Interactomes in Living Cells with Nanometer Resolution

在这个生物技术分子基础(MFB)项目中，斯坦福大学的Alice Ting博士和她的团队将致力于开发研究RNA的新技术，RNA是携带遗传信息、编码蛋白质并在细胞功能中发挥关键作用的基本分子。传统上，我们对RNA的理解一直受到需要分裂细胞的方法的限制，导致信息丢失。该团队的目标是创造非破坏性技术，使科学家能够直接研究活细胞中的RNA，为它们的位置、相互作用和功能提供新的见解。这项研究对人类健康特别重要，因为RNA的中断与包括癌症在内的许多疾病有关。该项目超越了它的科学影响，通过促进合作、举办研讨会和参与外联活动，将未被充分代表的学生介绍给令人兴奋的RNA科学世界。该项目通过利用先前用于蛋白质的成熟的邻近标记(PL)技术，解决了目前在活细胞内研究RNA的局限性。凭借十多年的PL经验，研究团队的目标是：(1)开发新的方法用于绘制活细胞中RNA定位的转录组图谱，通过改进的标记化学方法实现单细胞分辨率；(2)通过蛋白质工程和定向进化创建以RNA为中心的交互组图工具；以及(3)将这些技术应用于癌症生物学，与专家合作探索肿瘤与免疫细胞的相互作用，并绘制与癌症相关的特定RNA相互作用图谱。该项目的智力优势源于该团队的多学科专业知识，涵盖蛋白质和RNA工程、定向进化、化学生物学、计算分析和癌症生物学。更广泛的影响包括在癌症治疗中创造广泛适用的工具、教育机会和潜在的进步，促进公共健康的改善。该项目得到了数学和物理科学局的化学部和生物科学局的分子和细胞生物科学与系统和合成生物学分部的支持。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

In this Molecular Foundations for Biotechnology (MFB) project, Dr. Alice Ting and her team at Stanford University will embark on an effort to develop new technologies for studying RNA, the essential molecules that carry genetic information, encode proteins, and play crucial roles in cell functions. Traditionally, our understanding of RNA has been limited by methods that require breaking cells apart, leading to information loss. The team's goal is to create non-destructive technologies that allow scientists to directly study RNA in living cells, offering new insights into their location, interactions, and functions. This research is particularly significant for human health, as disruptions in RNA are implicated in many diseases, including cancer. The project extends beyond its scientific impact by fostering collaborations, conducting workshops, and engaging in outreach activities to introduce under-represented students to the exciting world of RNA science.This project addresses current limitations in studying RNA within living cells by leveraging proven proximity labeling (PL) technologies previously used for proteins. Drawing on over a decade of PL experience, the research team aims to: (1) develop new methods for transcriptome-wide mapping of RNA localization in living cells, achieving single-cell resolution with improved labeling chemistries; (2) create RNA-centric interactome mapping tools through protein engineering and directed evolution; and (3) apply these technologies to cancer biology, collaborating with experts to explore tumor-immune cell interactions and map specific cancer-associated RNA interactomes. The project's intellectual merit stems from the team's multidisciplinary expertise, spanning protein and RNA engineering, directed evolution, chemical biology, computational analysis, and cancer biology. Broader impacts include the creation of widely applicable tools, educational opportunities, and potential advancements in cancer therapeutics, contributing to improvements in public health.This project is supported by the Division of Chemistry in the Directorate for Mathematical and Physical Sciences, and by the Division of Molecular and Cellular Biosciences and Systems and Synthetic Biology cluster in the Directorate for Biological Sciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.