As discovery of cellular diversity in the brain accelerates, so does the need for tools that target cells based on multiple features. Here we developed Conditional Viral Expression by Ribozyme Guided Degradation (ConVERGD), an adeno-associated virus-based, single-construct, intersectional targeting strategy that combines a self-cleaving ribozyme with traditional FLEx switches to deliver molecular cargo to specific neuronal subtypes. ConVERGD offers benefits over existing intersectional expression platforms, such as expanded intersectional targeting with up to five recombinase-based features, accommodation of larger and more complex payloads and a vector that is easy to modify for rapid toolkit expansion. In the present report we employed ConVERGD to characterize an unexplored subpopulation of norepinephrine (NE)-producing neurons within the rodent locus coeruleus that co-express the endogenous opioid gene prodynorphin (Pdyn). These studies showcase ConVERGD as a versatile tool for targeting diverse cell types and reveal Pdyn-expressing NE⁺ locus coeruleus neurons as a small neuronal subpopulation capable of driving anxiogenic behavioral responses in rodents.

随着大脑细胞多样性的发现加速，对基于多种特征的针对细胞的工具的需求也在增加。在这里，我们开发了核酶引导降解条件病毒表达（ConVERGD），这是一种基于腺相关病毒的单一构建体交叉靶向策略，它将自切割核酶与传统的 FLEx 开关相结合，将分子货物传递到特定的神经元亚型。 ConVERGD 提供了优于现有交叉表达平台的优势，例如具有多达五个基于重组酶的功能的扩展交叉靶向、容纳更大和更复杂的有效负载以及易于修改以实现快速工具包扩展的载体。在本报告中，我们采用 ConVERGD 来表征啮齿动物蓝斑内未探索的产生去甲肾上腺素 (NE) 的神经元亚群，这些神经元共同表达内源性阿片类药物基因前强啡肽 ( Pdyn )。这些研究展示了 ConVERGD 作为针对不同细胞类型的多功能工具，并揭示了表达Pdyn的 NE ⁺蓝斑神经元是一种能够驱动啮齿类动物产生焦虑行为反应的小型神经元亚群。