Collaborative Research: Ideas Lab: RNA-encoded Molecular Memory (REMM)

合作研究：创意实验室：RNA 编码的分子记忆 (REMM)

• 批准号: 2243698
• 负责人: Saad Bhamla
• 金额: $ 5万
• 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: 2023-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2243698&HistoricalAwards=false
• 关键词: Collaborative; Research; Ideas; Lab; RNA

In most organisms, the genetic information is composed of the molecule deoxyribonucleic acid (DNA). But only a fraction of DNA in the genome—the “coding DNA”—contains biochemical instructions for generating the proteins that perform the vast range of activities essential for life. Indeed, 99% of the human genome is regarded as “non-coding” DNA. Intriguingly, the DNA in this “non-coding” part of the genome can still generate RNA, even though these non-coding RNAs are not translated into proteins, in contrast to the "messenger" RNAs generated by coding DNA. Surprisingly, the functions of non-coding RNAs, which represent a large percentage of the genome, are still largely unknown. A major goal of the present project is to explore potential roles these non-coding RNAs could play in the encoding of memory. At present, memories are widely thought to be stored as changes in the strengths of connections among neurons in the brain. The project will build on prior evidence that some forms of simple memory may instead be encoded as changes in RNA molecules. The project will examine whether one simple form of memory—behavioral sensitization—can be stored by non-coding RNA, or whether non-coding RNA can produce downstream molecular changes that store memory. In addition, the project will enhance the training of future scientists, including graduate students and postdoctoral researchers. Finally, undergraduate and minority students underrepresented in STEM will participate in the proposed research under the mentorship of the principal investigators, and thereby gain a deeper understanding of scientific research.A previous study by one of the principal investigators (PIs) showed that long-term memory (LTM) can be transferred by injecting RNA from trained animals into untrained animals. In this prior work, RNA was extracted from the nervous systems of sensitized (“trained”) marine snails (Aplysia californica), purified, and then injected into untrained snails; injected RNA from trained animals produced sensitization in untrained animals, whereas RNA from untrained donor animals did not. The PIs hypothesize that the expression or post-transcriptional state of one or more non-coding RNAs (ncRNAs) is selectively induced by sensitization training and that this molecular change mediates persistent sensitization memory in Aplysia. To test this hypothesis, the PIs will perform differential RNA-seq on purified RNA from trained and untrained animals and identify species of ncRNA with changes that correlate significantly with LTM. Initial experiments will determine whether the expression levels of specific ncRNAs change as a consequence of sensitization. Based on these results, additional aspects of ncRNA structure, modification state or protein interactions will be probed to explore whether sensitization training induces persistent changes in these ncRNA properties that could mediate memory encoding in Aplysia. This work would set the stage for a future project in which candidate ncRNAs be screened for mnemonic potency by disrupting them with antisense oligonucleotides (ASOs) and assessing the effects on neuronal excitability and synaptic connectivity of Aplysia sensory and motor neurons, as well as the effect of the ASOs on LTM in intact animals.This award was co-funded by the Directorate for Biological Sciences, and the Neural Systems Cluster in the Division of Integrative Organismal 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.

在大多数生物体中，遗传信息由分子脱氧核糖核酸（DNA）组成。但是，基因组中只有一小部分DNA（“编码DNA”）辅助生化指令，以产生执行对生命必不可少的活性的蛋白质。实际上，99％的人基因组被认为是“非编码” DNA。有趣的是，与编码DNA产生的“ Messenger” RNA相比，基因组的“非编码”部分中的DNA仍然可以产生RNA。令人惊讶的是，代表大部分基因组的非编码RNA的功能仍然很大程度上是未知的。本项目的主要目标是探索这些非编码RNA可以在内存编码中扮演的潜在角色。目前，人们普遍认为，记忆被认为是大脑神经元之间连接强度的变化。该项目将基于先前的证据，表明某些形式的简单记忆可能被编码为RNA分子的变化。该项目将检查一种简单的记忆形式（行为灵敏度）是否可以通过非编码RNA来存储，或者非编码RNA是否可以产生存储记忆的下游分子变化。此外，该项目将增强对未来科学家的培训，包括研究生和博士后研究人员。最后，本科生和少数族裔学生在STEM中的代表性不足将在主要研究人员的心态下参与拟议的研究，从而对科学研究有了更深入的了解。一项主要研究人员（PIS）先前的研究表明，长期记忆（LTM）可以通过将RNA从训练有素的动物中注入未经训练的动物中。在这项先前的工作中，从敏感（训练有素的）海洋蜗牛（加利福尼亚州阿普利西亚）的神经系统中提取RNA，纯化，然后注入未经训练的蜗牛中。受过训练的动物的注射RNA在未经训练的动物中产生敏感性，而未经训练的供体动物的RNA则没有。 PIS假设一个或多个非编码RNA（NCRNA）的表达或转录后态被选择性地通过灵敏度训练诱导，并且这种分子变化介导了Aplysia中的持续敏化记忆。为了检验这一假设，PI将对受过训练和未经训练的动物的纯化RNA执行差异RNA-Seq，并鉴定NCRNA的物种，其变化与LTM显着相关。初始实验将确定特定NCRNA的表达水平是否由于灵敏度而变化。基于这些结果，将研究NCRNA结构，修饰状态或蛋白质相互作用的其他方面，以探讨这些NCRNA特性中的灵敏度训练是否可以介导Aplysia中编码的记忆。这项工作将为未来的项目奠定阶段，在该项目中，通过使用反义寡核苷酸（ASO）破坏候选NCRNA，以筛选助记符，并评估对神经元的兴奋和突触连通性的影响，以及对LTM奖中的ASOBIND ASS COFUND的影响。该奖项反映了NSF的法定任务，并通过使用基金会的知识分子优点和更广泛的审查标准评估，反映了NSF的法定任务。