美国圣犹大儿童研究医院Lindsay A. Schwarz研究团队，报道了用于研究细胞多样性和大脑功能的单载体交叉AAV方法。2024年5月27日出版的《自然—神经科学》发表了这项成果。

据悉，随着大脑细胞多样性发现的加速，对靶向细胞多种特征工具的需求也在增加。

研究人员建立了一种基于腺相关病毒、单一结构的交叉靶向方法-由核酶引导降解的条件性病毒表达（ConVERGD）系统，其将自切割核酶与传统的FLEx开关结合起来，可将分子货物传递到特定的神经元亚群。与现有的交叉表达平台相比，ConVERGD具有很多优点，例如交叉靶向范围的扩大，具有多达五种重组酶的特征，可容纳更大、更复杂的有效载荷，而且载体易于修改，可快速扩展工具包。

研究人员利用ConVERGD揭示了啮齿动物小脑局灶内尚未发现的，去甲肾上腺素（NE）分泌神经元亚群的特征，这些神经元共同表达内源性阿片基因前吗啡（Pdyn）。这些研究表明， ConVERGD是一种针对不同细胞类型的通用工具，并揭示了表达NE⁺的啮齿动物局灶小叶神经元，是能够诱导啮齿动物产生焦虑反应的小神经元亚群。

附：英文原文

Title: A single-vector intersectional AAV strategy for interrogating cellular diversity and brain function

Author: Hughes, Alex C., Pittman, Brittany G., Xu, Beisi, Gammons, Jesse W., Webb, Charis M., Nolen, Hunter G., Chapman, Phillip, Bikoff, Jay B., Schwarz, Lindsay A.

Issue&Volume: 2024-05-27

Abstract: 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.

DOI: 10.1038/s41593-024-01659-7

Source: https://www.nature.com/articles/s41593-024-01659-7