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.

抽象的 大脑非常复杂，我们对该器官的理解仍处于起步阶段。许多 关于大脑发育和功能的基本问题仍然存在。单细胞基因组学承诺新方法 要回答这些问题，但是几乎所有的单细胞方法都有缺点 - 当细胞的细胞被破坏时 测量分子状态。这种“观察破坏”使得不可能将分子关联 过去发生的最终结果发生的事件：如果您摧毁了中间祖先 大脑要介绍其表观基因组，您无法知道祖细胞会采用哪种细胞命运。 同样，经历后对动物进行分析将使动物混合在一起的分子后果 在暴露前有任何分子倾向的经验。如果您在 经验，您无法了解动物的结果，例如如果动物将是学习者或 不是。这种缺点是继承的几乎所有现有基因组方法，并阻碍了各种各样的兴趣 分析。我们最近开发了一种平台技术，即使用的单细胞“电话卡”（CC） 转座子以一瞬间的时间捕获分子事件，然后以后再读取它们 细胞分辨率。现在，我们建议建立在这个基础上，以发展强大，易于使用的技术 在小鼠大脑中单细胞分辨率下记录分子事件的平台，并与细胞命运相连 决策，行为结果，神经元活动谱和解剖位置。我们将广泛开发一组 适用的试剂和方法，将它们应用于一系列高影响的“测试用例”以证明其实用性， 并迅速将它们分发给更广泛的社区。