Tools4Cells: EAGER: A Molecular Pursuit for the Engram: Microfluidic temporal transcriptomics for single cell learning

Tools4Cells：EAGER：对印迹的分子追求：用于单细胞学习的微流控时间转录组学

• 批准号: 2337788
• 负责人: Saad Bhamla
• 金额: $ 30万
• 依托单位: Georgia Tech Research Corporation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-11-01 至 2025-10-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2337788&HistoricalAwards=false
• 关键词: Tools4Cells; EAGER; Molecular; Pursuit; Engram

This research aims to elucidate the mechanisms by which RNA encodes memory and learning in single cells. Despite their lack of neurons, single-celled organisms, such as Spirostomum ambiguum, demonstrate non-associative forms of learning, including habituation. Intriguingly, these memories can be transferred between organisms, with accumulating evidence suggesting RNA as a 'memory molecule'. By leveraging the simplicity of these single- celled organisms, the project will explore behavioral aspects at the molecular level, bypassing the complexities of intercellular interactions. Through the development of innovative microfluidic temporal single-cell transcriptomic assays, the role of non-coding RNA (ncRNA) in learning and memory storage will be investigated. The project offers a multidisciplinary learning opportunity for a diverse range of graduate student scientists and scholars at different stages in their academic careers. The project will also develop bilingual outreach comics to disseminate complex scientific ideas to a broader audience, fostering a deeper understanding of the research and its implications. The proposed research could validate RNA as a molecular memory, enabling long-term information storage and transfer in organisms. Studies have shown that RNA holds information that can transfer memories between organisms in multicellular species like C. elegans, Aplysia, flatworms, rats, and fish, challenging a purely synaptic view. To consider potential applications of the phenomena, we must first prove that a molecular engram such as RNA contains information that can mediate behavior. To decipher the potential information encoded within RNA, this EAGER will: i) develop appropriate microfluidic hardware that allows for non-destructive RNA extraction, ii) optimize the workflow and sequencing protocol to create a low-input RNA-seq approach for reliable live cell transcriptomics, and iii) combine these elements to monitor RNA changes, speciation, and spatiotemporal expression profiles during habituation and memory transfer. The results could significantly change our understanding of memory processing, testing the capacity of information-encoded RNA substrates to be stored, modified, and even transferred between living and synthetic systems.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.

这项研究旨在阐明RNA编码单个细胞中记忆和学习的机制。尽管缺乏神经元，但单细胞的生物（例如螺旋体狂热）表现出非缔合性的学习形式，包括习惯。有趣的是，这些记忆可以在生物体之间传递，积累的证据表明RNA是“记忆分子”。通过利用这些单细胞生物的简单性，该项目将探索分子水平的行为方面，绕过细胞间相互作用的复杂性。通过开发创新的微流体单细胞转录组分析，将研究非编码RNA（NCRNA）在学习和记忆存储中的作用。该项目为在学术职业的不同阶段提供多种研究生科学家和学者提供了多学科学习的机会。该项目还将开发双语外展漫画，以向更广泛的受众传播复杂的科学思想，从而深入了解研究及其含义。拟议的研究可以将RNA验证为分子记忆，从而实现生物体中的长期信息存储和转移。研究表明，RNA拥有可以在秀丽隐杆线虫，aplysia，lind虫，大鼠和鱼类等多细胞物种中传递记忆的信息，挑战了纯粹的突触视图。为了考虑现象的潜在应用，我们必须首先证明RNA之类的分子ENGRAM包含可以介导行为的信息。为了解释RNA中编码的潜在信息，此急切将：i）开发适当的微流体硬件，允许非破坏性RNA提取，ii）优化工作流程和测序协议，以创建低输入RNA-SEQ方法，以创建可靠的活细胞转录组学，以及在RNA的习惯中结合这些元素的习性和SPATIOTEM，SPATIOTEM，SPATIOTEM，SPATIOTEM，SPATIOTEM和SPATATEMEM CRIPTION CLAIMETIOM，ii）。结果可能会大大改变我们对记忆处理的理解，测试信息编码的RNA底物的能力，以存储，修改，甚至在生活和合成系统之间进行转移。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛的影响审查的审查标准来通过评估来通过评估来获得支持的。