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

MFB：下一代邻近标记技术以纳米分辨率绘制活细胞中的亚细胞转录组和 RNA 相互作用组图

• 批准号: 2330686
• 负责人: Alice Ting
• 金额: $ 150万
• 依托单位: Stanford University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2330686&HistoricalAwards=false
• 关键词: MFB; Next; generation; Proximity; Labeling

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.

在这个生物技术分子基础 (MFB) 项目中，Alice Ting 博士和她在斯坦福大学的团队将致力于开发研究 RNA 的新技术，RNA 是携带遗传信息、编码蛋白质并在生物多样性中发挥关键作用的重要分子。细胞功能。传统上，我们对 RNA 的理解一直受到需要将细胞分开的方法的限制，从而导致信息丢失。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。这项研究对于人类健康尤其重要，因为 RNA 的破坏与包括癌症在内的许多疾病有关。该项目通过促进合作、举办研讨会和参与外展活动，向代表性不足的学生介绍令人兴奋的 RNA 科学世界，从而超越了其科学影响。该项目利用经过验证的邻近标记解决了当前在活细胞内研究 RNA 方面的局限性（ PL）技术以前用于蛋白质。凭借十多年的 PL 经验，研究团队的目标是：（1）开发用于活细胞中 RNA 定位全转录组图谱的新方法，通过改进的标记化学实现单细胞分辨率； (2)通过蛋白质工程和定向进化创建以RNA为中心的相互作用组图谱工具； (3) 将这些技术应用于癌症生物学，与专家合作探索肿瘤-免疫细胞相互作用并绘制特定的癌症相关 RNA 相互作用组图。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。更广泛的影响包括创建广泛适用的工具、教育机会和癌症治疗的潜在进步，从而有助于改善公共卫生。该项目得到了数学和物理科学理事会化学部以及生物科学理事会的分子和细胞生物科学以及系统和合成生物学集群。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。