Matthews Elizabeth A, Russ Jeffrey B, Qian Yongjun, Zhao Shengli, Thompson Peyton, Methani Muhib, Vestal Matthew, Huang Z Josh, Southwell Derek
Department of Neurosurgery, Duke University, Durham, NC, USA; Department of Neurobiology, Duke University, Durham, NC, USA.
Department of Neurosurgery, Duke University, Durham, NC, USA; Department of Pediatrics, Duke University, Durham, NC, USA.
Cell Rep. 2025 Aug 26;44(8):116037. doi: 10.1016/j.celrep.2025.116037. Epub 2025 Jul 22.
Reliable and systematic access to diverse cell types is necessary for understanding the organization, function, and pathophysiology of human neural circuits. Methods for targeting human neural populations are scarce and currently center on identifying transcriptional enhancers and engineering viral capsids. Here, we demonstrate the utility of cell access through RNA sensing by endogenous adenosine deaminase acting on RNA (ADAR) (CellREADR), a programmable RNA sensor-effector technology that couples cellular RNA sensing to effector protein translation, for accessing, monitoring, and manipulating specific neuron types in the human cortex ex vivo. We design CellREADRs to target two subpopulations-calretinin (CALB2) GABAergic interneurons and forkhead box protein P2 (FOXP2) glutamatergic projection neurons-and then validate targeting specificity using histological, electrophysiological, and transcriptomic methods. CellREADR expression of channelrhodopsin and GCamp enables the manipulation and monitoring of these populations in live cortical microcircuits. By demonstrating specific, reliable, and programmable experimental access to human neuronal subpopulations, our results highlight CellREADR's potential for studying neural circuits and treating brain disorders.
可靠且系统地获取多种细胞类型对于理解人类神经回路的组织、功能和病理生理学至关重要。靶向人类神经群体的方法稀缺，目前主要集中在识别转录增强子和改造病毒衣壳。在这里，我们展示了通过作用于RNA的内源性腺苷脱氨酶（ADAR）进行RNA传感来实现细胞访问的实用性（CellREADR），这是一种可编程的RNA传感器效应技术，可将细胞RNA传感与效应蛋白翻译偶联起来，用于在体外访问、监测和操纵人类皮质中的特定神经元类型。我们设计CellREADR以靶向两个亚群——钙视网膜蛋白（CALB2）γ-氨基丁酸能中间神经元和叉头框蛋白P2（FOXP2）谷氨酸能投射神经元——然后使用组织学、电生理学和转录组学方法验证靶向特异性。通道视紫红质和GCamp的CellREADR表达能够在活体皮质微回路中操纵和监测这些群体。通过展示对人类神经元亚群的特异性、可靠且可编程的实验访问，我们的结果突出了CellREADR在研究神经回路和治疗脑部疾病方面的潜力。
