Wyoming Sensory Biology COBRE

这份行政副刊的目的是提高我们在转录方面的研究能力。 神经系统的特征，并通过以下方式在SBC调查人员之间建立更协同的互动 促进转录组研究和合作。这些资金将用于收购NextSeq550 测序系统(Illumina)，这将大大加强单细胞克隆获得转录转录研究的机会，以及 加强集成显微镜核心的分子分析单元(MAU)的服务 (IMCore)。在过去4年的筹资期间，IMCore在援助方面取得重大进展 SBC项目负责人与他们的研究涉及转录组。目前，已经有了单细胞 RNAseq和Chipseq实验已经在核心进行，这在我们最近的出版物中得到了证明。 然而，这些实验只能通过将样本运送给多个三分之一的人来进行。 派对服务中心和核心设施(例如，Genewiz.com 10x基因组学和哈佛鲍尔核心设施)。 虽然这种方法已经足够了，但有几个主要问题阻碍了它在SBC研究人员中的应用：1) 运送的样品需要零下80华氏度/摄氏度的存储，用干冰运输它们会增加 样品质量的不必要变化；2)图书馆制剂的质量控制高度可变 因为它们是由使用不同协议的不同商业供应商进行的；3)缺乏实践技术 支持会阻止新项目负责人的申请。因此，购买NextSeq550将是一场游戏 怀俄明大学转录学研究的更改者。我们预计，项目的数量 使用单细胞RNAseq或Chipseq方法将显著增加，因为 NextSeq550。收购NextSeq550是完全合理的，合理的，可分配给授予项目-我们 目前有四个新的研究项目概述了使用该仪器的情况。项目1：确定国家-- 在单个细胞水平上神经元转录的依赖性变化。项目2：单核RNA-SEQ 将被用来探索具有差异投影的内侧前额叶皮质的转录异质性 在压力导致的抑郁期间。项目3：研究特定奖励群体中的基因表达-- 定义为同步激活的离散数量的神经元-以及这在神经元中的差异 在寻求不同类型的奖励的过程中被激活。项目4：回答以下问题 问题：(1)哪些基因在损伤后和再生过程中被激活/抑制，以及(2) 不同的损伤机制和/或细胞类型产生不同的转录图谱。我们预计这将是 行政补充将大大有助于增加我们获得转录研究和 协作，增加在剩余时间内使用此工具的项目数量 资助期，并有助于为第二阶段的申请奠定坚实的基础。

The objective of this administrative supplement is to enhance our research capabilities in the transcriptomic characterization of the nervous system, and forge a more synergistic interaction among SBC investigators by promoting transcriptomic research and collaborations. These funds will be used to acquire a NextSeq550 Sequencing System (Illumina) that will significantly enhance SBC’s access to transcriptomic research, and strengthen the services provided by the Molecular Analysis Unit (MAU) of the Integrated Microscopy Core (IMCore). Over the past 4-year funding period, the IMCore has made significant advances towards assisting SBC project leaders with their research involving transcriptomics. Currently, there have already been single cell RNAseq and Chipseq experiments that have been conducted in the core, as evidenced in our recent publications. However, these experiments were only able to be carried out through shipping out samples to multiple third- party service centers and core facilities (e.g., Genewiz.com 10x genomics and Harvard Bauer Core Facilities). Although this approach has “sufficed”, several major issues hinder its application among SBC researchers: 1) the shipped samples require -80 degree Fahrenheit/Celsius storage, and transporting them in dry ice adds unnecessary variability to the sample qualities; 2) the quality controls of library preparations are highly variable as they are conducted by different commercial vendors that use different protocols; 3) lack of hands-on technical support inhibits application by new project leaders. Therefore, the purchase of the NextSeq550 will be a game changer for transcriptomic research at the University of Wyoming. We anticipate that the number of projects using single cell RNAseq or the Chipseq approach will increase significantly given the availability of the NextSeq550. The purchase of NextSeq550 is fully justifiable, reasonable, and allocable to grant projects – we currently have four new research projects that outline utilizing this instrument. Project 1: to identify state- dependent changes in neuronal transcription at the level of individual cells. Project 2: single-nucleus RNA-seq will be used to explore transcriptional heterogeneity in the medial prefrontal cortex with differential projections during stress-induced depression. Project 3: to investigate gene expression within reward-specific ensembles – defined as a discrete number of neurons that become synchronously activated – and how that differs in neurons that become activated during the seeking of different types of rewards. Project 4: to answer the following questions: (1) which genes are activated/repressed after injury and during regeneration, and (2) are the transcriptional profiles generated different based on injury mechanism and/or cell type. We anticipate that this administrative supplement will significantly help increase our access to transcriptomic research and collaboration, increase the number of projects that would utilize this instrument during the remainder of the funding period, and help build a strong foundation for Phase II applications.