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， 这对于实现有针对性的电路操作以了解正常的大脑功能和大脑至关重要。使用 靶向基因表达的基因调控元件正在改变脑回路研究。响应于 RFA-MH-21-180，神经回路映射中心（CNCM）团队，由少数民族大学徐祥民博士领导。 服务机构（MSI）-指定机构，加州大学欧文分校（UCI）将与博士合作。 戈登·菲谢尔的团队在哈佛大学和布罗德研究所生产和销售的试剂开发 通过他们的Armamentarium项目，“系统识别增强剂，以靶向兴奋性和 大脑皮层中的抑制性神经元细胞类型”（U 01 MH 13070，待裁决）。费谢尔博士的团队 建立了一个新的高通量增强子筛选平台，使用基因调控元件， 皮层GABA能中间神经元和锥体兴奋性神经元亚型的细胞类型限制性基因表达 前所未有的决议。这是通过使用腺相关病毒（AAV）载体来实现的， 筛选和验证小鼠、非人灵长类动物和人中特定神经元类型的增强子 个脑袋我们提出的研究的第一个目标是建立UCI和哈佛之间的密切合作 大学/布罗德研究所团队扩大和优化细胞类型特异性增强子-AAV的生产。 利用我们不断发展的中心平台，UCI团队将把这些病毒试剂分发给合格的研究人员， 神经科学界对细胞类型特异性的访问和操作。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， 为少数民族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Fishell博士的团队一起扩大Fishell博士团队的种子增强剂-AAV试剂， 一套广泛的工具，用于脊椎动物物种的细胞类型特异性神经回路研究。我们将利用我们 CNCM现有的嗜神经病毒生产和分销的专业知识，使新的辅助AAV的基础上博士。 Fishell的增强子，以提高CNS中遗传靶向特异性电路映射的精度。目标 2和3，我们将与Fishell博士的试点U 01项目研究人员协调，以扩大我们的UCI CNCM病毒 生产线，并改善我们的分销平台，以支持病毒试剂的生产，验证， 为研究社区分配充足的物资。我们将扩大我们的团队成员的招聘， 通过与西部健康科学大学和莫尔豪斯学校合作，促进了不同的背景 医学包括教师，工作人员和学生的代表性不足的少数民族的拟议研究。我们 将满足RFA要求，以规范的方式生产和分发经认证的病毒试剂 被广泛应用于研究领域。这个U24奖项将为BRAIN倡议提供关键支持 军械库项目。

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.