UC Irvine Center for the production and distribution of cell-type-specific viral targeting reagents

项目摘要 在啮齿动物、非人类灵长类动物和其他脊椎动物物种中获得特定细胞类型的遗传途径是 对于使有针对性的电路操作能够了解正常的大脑功能和大脑至关重要。对.的使用 靶向基因表达的基因调控元件正在改变大脑电路的研究。作为对.的回应 RFA-MH-21-180，少数民族徐向民博士领导的神经回路标测中心(CNCM)团队 服务机构(MSI)-指定机构，加州大学欧文分校(UCI)将与Dr。 戈登·菲舍尔的团队在哈佛大学和布罗德研究所生产和分销开发的试剂 通过他们的医疗设备项目，“系统地识别增强剂，以靶向兴奋性和 大脑皮层中的抑制性神经细胞类型“(U01MH13070，待奖)。菲舍尔博士的团队已经 利用基因调控元件建立了一种新型的高通量增强子筛选平台，使 皮质GABA能中间神经元和锥体兴奋性神经元亚型细胞类型限制性基因的表达 以前所未有的决心。这是使用腺相关病毒(AAV)载体实现的有效 小鼠、非人灵长类和人类特定神经元类型增强剂的筛选和验证 大脑。我们提出的研究的第一个目标是在UCI和哈佛之间建立密切的合作 大学/博德研究所团队扩大和优化特定细胞类型的增强子-AAVs的生产。 使用我们的成长中心平台，UCI团队将在年将这些病毒试剂分发给合格的调查人员 神经科学社区提供特定细胞类型的访问和操作。我们研究的第二个目标是 通过吸引来自不同背景的科学家和学生来增强该项目的影响力 为少数群体社区服务的研究密集型机构。在目标1中，UCI团队将建立合作伙伴关系 与菲舍尔博士的团队一起扩展种子增强剂-AAV试剂-菲舍尔博士的团队所表征的 一套广泛的工具，用于研究脊椎动物物种中特定细胞类型的神经回路。我们将利用我们的 CNCM在神经性病毒的生产和分发方面拥有现有的专业知识，以DR为基础制造新的辅助AAVs。 FISHELL的增强剂，以提高中枢神经系统中基因靶向特定电路映射的精确度。在AIMS中 2和3，我们将与菲舍尔博士的U01试点项目调查人员协调，扩大我们的UCI CNCM病毒 生产流水线和改进我们的分销平台，以支持病毒试剂的生产、验证和 为研究社区分发提供充足的供应。我们将扩大对团队成员的招聘 与西方健康科学大学和莫尔豪斯学院合作促进不同背景 医学部将少数族裔的教职员工和学生纳入拟议的研究。我们 将满足RFA的要求，以规范的方式生成和分发经过验证的病毒试剂 在研究界得到广泛应用。这个U24奖项将为Brain-Initiative提供关键支持 兵器馆项目。

Project Summary Gaining genetic access to specific cell types in rodents, non-human primates and other vertebrate species is critical for enabling targeted circuit manipulations to understand normal brain function and brain. The use of gene regulatory elements for targeted gene expression is transforming brain circuitry studies. In response to RFA-MH-21-180, the Center for Neural Circuit Mapping (CNCM) team led by Dr. Xiangmin Xu at the Minority Serving Institution (MSI)-designated institution, University of California, Irvine (UCI) will collaborate with Dr. Gordon Fishell’s team at Harvard University and the Broad Institute to produce and distribute reagents developed by their Armamentarium project, “Systematic identification of enhancers to target the breadth of excitatory and inhibitory neuronal cell types in the cerebral cortex” (U01MH13070, pending award). Dr. Fishell’s team has established a novel high-throughput enhancer screening platform using gene regulatory elements that enables cell type-restricted gene expression in cortical GABAergic interneuron and pyramidal excitatory neuron subtypes at an unprecedented resolution. This is achieved using adeno-associated virus (AAV) vectors for the effective screening and validation of enhancers for specific neuron types in the mouse, non-human primate, and human brain. The first goal of our proposed research is to establish close collaborations between the UCI and Harvard University/Broad Institute teams to scale up and optimize the production of cell-type-specific enhancer-AAVs. Using our growing Center platform, the UCI team will distribute these viral reagents to qualified investigators in the neuroscience community for cell-type-specific access and manipulation. The second goal of our research is to enhance the impact of this project by engaging scientists and students from diverse backgrounds and less research-intensive institutions that serve minority communities. In Aim 1, the UCI team will form a partnership with Dr. Fishell’s team to expand seed enhancer-AAV reagents characterized by Dr. Fishell’s team to make a broad set of tools for cell-type-specific neural circuit studies across vertebrate species. We will leverage our CNCM existing expertise in neurotropic virus production and distribution to make new helper AAVs based on Dr. Fishell’s enhancers to improve the precision of genetically targeted specific circuit mapping in the CNS. In Aims 2 and 3, we will coordinate with Dr. Fishell’s pilot U01 project investigators to scale up our UCI CNCM viral production pipeline and improve our distribution platform to support viral reagent production, validation, and ample supplies for research community distribution. We will expand our recruitment of team members from diverse backgrounds facilitated by partnering with Western University of Health Sciences and Morehouse School of Medicine to include faculty, staff and students of underrepresented minorities for the proposed research. We will fulfill the RFA requirement to generate and distribute authenticated viral reagents in a well-regulated manner to be widely used in the research community. This U24 award will provide critical support for the BRAIN-Initiative Armamentarium project.