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

项目总结 神经科学中的一个根本挑战是了解大脑中的哪些回路和细胞对 特定的行为、感知和功能。一类工具，可以帮助揭示复杂的大脑电路 是活动集成器，可以记录在用户指定的时间窗口内神经活动的历史。 技术不仅记录和映射神经活动，而且使基因能够获得“回放”神经活动 可以提供对大脑回路结构的无与伦比的了解。该项目将开发和验证新的 光遗传工具，允许在一定时间内对体内最近激活的神经元进行永久遗传访问 用光指定的。该项目建立在先前开发和验证的光遗传工具基础上：a 一种背景低、诱导活性高、范围窄的可光激活DNA重组酶技术 激活的时间窗口。我们将在培养细胞和小鼠脑内验证这些工具。有了这些 工具，用户可以在行为过程中施加短暂的光脉冲，导致永久的基因修改 在行为过程中，神经元处于活跃状态。这些精密工具将为神经科学家和 使各种新的研究能够在正常和疾病中询问和操纵复杂的大脑电路 大脑。

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.