Molecular recording to predict cell fate decisions and animal behavior

分子记录预测细胞命运决定和动物行为

• 批准号: 10260139
• 负责人: JOSEPH D DOUGHERTY
• 金额: $ 369.33万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10260139
• 关键词: Address; Adopted; Anatomy; Animal Behavior; Animals; Behavioral; Benchmarking; Binding; Brain; Brain region; Cells; ChIP-seq; Chemicals; Communities; Complex; Development; Dose; Embryo; Enhancers; Epigenetic Process; Esthesia; Event; Exposure to; Foundations; GABA Antagonists; Gene Expression; Genetic Transcription; Genome; Genomics; Grant; Inbred Mouse; Label; Lead; Learning; Link; Location; Measures; Mediating; Methods; Molecular; Mus; Neurons; Neurosciences; Organ; Organism; Outcome; Pain; Population; Positioning Attribute; Predisposition; Reagent; Resolution; Retina; Retinal Ganglion Cells; Seizures; Series; Spinal Ganglia; Stimulus; System; Techniques; Technology; Testing; Time; Training; Transposase; Viral; Whole Organism; behavior influence; behavioral outcome; cell type; conditioned fear; epigenome; experience; genome-wide; infancy; interest; jun Oncogene; learning outcome; mRNA Expression; nervous system disorder; postnatal; prevent; progenitor; response; social; social learning; transcription factor; transcriptome sequencing; transcriptomics

Abstract The brain is remarkably complex, and our understanding of this organ is still in its infancy. Many fundamental questions about brain development and function remain. Single cell genomics promises new ways to answer these questions, but nearly all single-cell methods share shortcoming – cells are destroyed when their molecular states are measured. This “destruction upon observation” makes it impossible to correlate molecular events that have occurred in the past with the final outcomes: if you destroy an intermediate progenitor in the brain to profile its epigenome, you have no way to know what cell fate the progenitor would have adopted. Likewise, profiling an animal after an experience is going to conflate the molecular consequences of the experience with any molecular predispositions that were there before exposure. And if you profile before the experience, you have no way of know of the animal's outcome – e.g. if the animal would have been a learner or not. This shortcoming is inherent to nearly all existing genomic methods and impedes a wide variety of interesting analyses. We have recently developed a platform technology, single-cell `Calling Cards' (CC) that uses transposons to capture molecular events at one instant in time and then read them out at a later time with single cell resolution. We now propose to build upon this foundation to develop a robust, easy to use technology platform to record molecular events at single cell resolution in the mouse brain and connect them with cell fate decision, behavioral outcomes, neuronal activity profiles, and anatomical location. We will develop a set of widely applicable reagents and methods, apply them to a series of high-impact “test cases” to demonstrate their utility, and rapidly distribute them to the broader community.

摘要 大脑非常复杂，我们对这个器官的了解还处于初级阶段。许多 关于大脑发育和功能的基本问题仍然存在。单细胞基因组学有望找到新的方法 来回答这些问题，但几乎所有的单细胞方法都有缺点--当细胞的 测量分子的状态。这种“一看就破坏”的现象使得人们不可能将分子 过去发生的具有最终结果的事件：如果您在 大脑要描绘它的表观基因组，你无法知道祖先会采用什么样的细胞命运。 同样，在经历过一次之后描绘一只动物将会混淆 在暴露前就有过任何分子易感性的经验。如果你在这之前分析 经验，你无法知道动物的结果--例如，如果动物是学习者或 不。这一缺陷是几乎所有现有基因组方法所固有的，并阻碍了各种各样有趣的 分析。我们最近开发了一种平台技术，单蜂窝呼叫卡(CC)，它使用 转座子在某个时刻捕捉分子事件，然后在稍后的时间用Single 单元格分辨率。我们现在建议在此基础上开发一种强大、易于使用的技术 记录小鼠大脑中单细胞分辨率的分子事件并将其与细胞命运联系起来的平台 决策、行为结果、神经元活动特征和解剖位置。我们将开发一套广泛的 适用的试剂和方法，将它们应用于一系列影响较大的“测试案例”，以证明它们的实用性， 并迅速将它们分发给更广泛的社区。