Yang Lite, Liu Fang, Hahm Hannah, Okuda Takao, Li Xiaoyue, Zhang Yufen, Kalyanaraman Vani, Heitmeier Monique R, Samineni Vijay K
Department of Anesthesiology, Washington University Pain Center, Washington University School of Medicine, St. Louis, MO, USA.
Neuroscience Graduate Program, Division of Biology & Biomedical Sciences, Washington University School of Medicine, St. Louis, MO, USA.
Nat Commun. 2025 Jul 1;16(1):5557. doi: 10.1038/s41467-025-60360-w.
Single-cell multiomic techniques have sparked immense interest in developing a comprehensive multi-modal map of diverse neuronal cell types and their brain-wide projections. However, investigating the complex wiring diagram, spatial organization, transcriptional, and epigenetic landscapes of brain-wide projection neurons is hampered by the lack of efficient and easily adoptable tools. Here we introduce Projection-TAGs, a retrograde AAV platform that allows multiplex tagging of projection neurons using RNA barcodes. By using Projection-TAGs, we performed multiplex projection tracing of the cortex and high-throughput single-cell profiling of the transcriptional and epigenetic landscapes of the cortical projection neurons in female mice. Projection-TAGs can be leveraged to obtain a snapshot of activity-dependent recruitment of distinct projection neurons and their molecular features in the context of a specific stimulus. Given its flexibility, usability, and compatibility, we envision that Projection-TAGs can be readily applied to build a comprehensive multi-modal map of brain neuronal cell types and their projections.
单细胞多组学技术引发了人们对绘制各种神经元细胞类型及其全脑投射的全面多模态图谱的浓厚兴趣。然而，由于缺乏高效且易于采用的工具，对全脑投射神经元的复杂布线图、空间组织、转录和表观遗传景观的研究受到了阻碍。在这里，我们介绍了Projection-TAGs，这是一种逆行腺相关病毒（AAV）平台，它允许使用RNA条形码对投射神经元进行多重标记。通过使用Projection-TAGs，我们对雌性小鼠的皮质进行了多重投射追踪，并对皮质投射神经元的转录和表观遗传景观进行了高通量单细胞分析。Projection-TAGs可用于获取特定刺激背景下不同投射神经元的活动依赖性募集及其分子特征的快照。鉴于其灵活性、可用性和兼容性，我们设想Projection-TAGs可以很容易地应用于构建大脑神经元细胞类型及其投射的全面多模态图谱。
