Precision editing of neural circuits using engineered electrical synapses

标题：使用工程化的电子突触精确编辑神经电路 包括光遗传学和设计者受体在内的开创性方法 设计药物(DREADD)能够从基因上直接调节个体的活动 定义的单元格类型。尽管如此，有选择性地对 神经回路的显著特征：两个特定脑细胞之间的接口。要解决这个问题 挑战，我们创造了一种新的方法，使用连接蛋白长期整合电路 (LinCx)，它使用一对新型的工程连接蛋白半通道来直接调制 由基因定义的神经回路。当半通道对的每个成员以 两个不同的单元格(S)/单元格类型组成一个电路，他们进行异型对接 (相互对接)，在两个细胞之间构成了一个电子突触。这些 一对半通道的设计是为了防止它们进行同型对接 (与自己形成电突触)，以及2)扰乱它们与他人的对接 连接蛋白半突触在哺乳动物的脑中内源性表达。最后，3) 半通道对表现出整形。这三个属性一起赋予LinCx 前所未有的空间、时间和上下文精确度，支持对神经网络的精确编辑 电路。 我们建议在模型生物中部署LinCx。我们将确定LinCx的影响 神经调节对神经回路生理和情绪行为的影响。我们还将测试 LinCx的调节足以恢复精神分裂症动物模型的正常行为 精神错乱。这些高风险实验的成功完成将产生一种新的方法 临床前模型中调节情绪状态的长期回路编辑。在未来，LinCx 可以与新出现的病毒工具集成，从而实现系统地传递基因编码的 特定脑细胞类型的蛋白质。因此，LinCx也有一条通向人类的道路 用于改善破坏性精神障碍的翻译。

Title: Precision editing of neural circuits using engineered electrical synapses Pioneering approaches including optogenetics and designer receptors exclusively activated by designer drugs (DREADDs) enable the direct modulation of the activity of individual genetically defined cell types. Nevertheless, it remains a fundamental challenge to selectively regulate the hallmark feature of neural circuits: the interface between two specific brain cells. To address this challenge, we have created a new approach, Long-term integration of circuits using Connexins (LinCx), that employs a novel pair of engineered connexin hemichannels to directly modulate genetically defined neural circuits. When each member of the hemichannel pair is expressed in two different cell(s)/cell-types that compose a circuit, they engage in heterotypic docking (docking with each other) and an electrical synapse is constituted between the two cells. These pair of hemichannels is engineered 1) to prevent them from engaging in homotypic docking (forming electrical synapses with themselves), and 2) to disrupt them from docking with other connexin hemichannels endogenously expressed in the mammalian brain. Finally, 3) the hemichannel pair exhibits rectification. Together, these three properties confer LinCx with unprecedented spatial-, temporal-, and context precision, enabling the precise editing of neural circuits. We propose to deploy LinCx across model organisms. We will determine the impact of LinCx neuromodulation on neural circuit physiology and emotional behavior. We will also test whether LinCx modulation is sufficient to restore normal behavior in animal models of psychiatric disorders. Successful completion of these high-risk experiments will yield a new method for long-term circuit editing to regulate emotional states in preclinical models. In the future, LinCx can be integrated with emerging viral tools that enable systemic delivery of genetically encoded proteins to specific brain cell-types. Thus, LinCx also has an attainable path to human translation for ameliorating devastating psychiatric disorders.