Light-Induced Targeted Recombination Strategies For Genetic Access to Recently Active Neurons

光诱导靶向重组策略对最近活跃的神经元进行遗传访问

• 批准号: 9924856
• 负责人: Aaron J Norris
• 金额: $ 48.4万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9924856
• 关键词: Adoption; Area; Behavior; Brain; Brain Diseases; Cells; Chemicals; Complex; Cultured Cells; DNA; Development; Disease; Doxycycline; Engineering; Enterobacteria phage P1 Cre recombinase; FOS gene; Fire - disasters; Gene Expression; Genetic; Genetic Recombination; Goals; Hour; In Vitro; Label; Light; Maps; Mediating; Memory; Methods; Modification; Molecular; Mus; Neurons; Neurosciences; Nuclear Import; Perception; Physiologic pulse; Plasmids; Population; Process; Proteins; Reactive Inhibition; Recording of previous events; Research Personnel; Resolution; Resources; Role; Specific qualifier value; Specificity; Stimulus; Structure; Swimming; System; Tamoxifen; Technology; Testing; Time; Transcription Coactivator; Validation; Visualization; Work; base; behavioral response; brain circuitry; design; experience; genetic approach; in vitro activity; in vivo; innovation; natural hypothermia; nervous system disorder; neural circuit; neuronal circuitry; novel; optogenetics; practical application; prevent; promoter; protein activation; recombinase; relating to nervous system; response; sensor; temporal measurement; time interval; tool; tool development

PROJECT SUMMARY A fundamental challenge in neuroscience is in understanding which circuits and cells in the brain contribute to specific behaviors, perceptions, and functions. One class of tools that can help to unveil complex brain circuitry are activity integrators that can record the history of neural activity over a user-specified window of time. Technologies that not only record and map neural activity but enable genetic access to ‘playback’ neural activity can provide an unparalleled understanding of brain circuit structures. This project will develop and validate new optogenetic tools allowing permanent genetic access to recently activated neurons in vivo, within a time frame specified using light. This project builds on a previously developed and validated optogenetic tool: a photoactivatable DNA recombinase technology that shows low background, high induced activity, and a narrow time window for activation. We will validate these tools in cultured cells and in vivo in mouse brain. With these tools, users could apply a brief pulse of light during a behavior, resulting in a permanent genetic modification in neurons active during the behavior. These precision tools will extend the genetic toolkit for neuroscientists and enable a variety of new studies interrogating and manipulating complex brain circuitry in the normal and diseased brain.

项目摘要 神经科学的一个基本挑战是了解大脑中的哪些回路和细胞有助于 具体的行为、感知和功能。有一类工具可以帮助揭示复杂的大脑回路 是活动积分器，可以记录用户指定时间窗口内的神经活动历史。 技术不仅记录和映射神经活动，而且使基因能够“回放”神经活动 可以提供对大脑回路结构的无与伦比的理解。该项目将开发和验证新的 光遗传学工具允许在一定时间范围内永久遗传进入体内最近激活的神经元 使用灯光指定。该项目建立在以前开发和验证的光遗传学工具上： 光活化DNA重组酶技术，显示出低背景、高诱导活性和窄的 激活的时间窗口。我们将在培养的细胞和小鼠大脑中验证这些工具。与这些 使用者可以在一个行为过程中施加一个短暂的光脉冲，导致永久的基因修改， 神经元在行为过程中活跃。这些精确的工具将扩展神经科学家的遗传工具包， 使各种新的研究能够询问和操纵正常和患病的复杂大脑回路， 个脑袋