Projection-TAGs enable multiplex projection tracing and multi-modal profiling of projection neurons

Lite Yang, Fang Liu, Hannah Hahm, Takao Okuda, Xiaoyue Li, Yufen Zhang, Vani Kalyanaraman, Monique R. Heitmeier and Vijay K. Samineni ()
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Lite Yang: Washington University School of Medicine
Fang Liu: Washington University School of Medicine
Hannah Hahm: Washington University School of Medicine
Takao Okuda: Washington University School of Medicine
Xiaoyue Li: Washington University School of Medicine
Yufen Zhang: Washington University School of Medicine
Vani Kalyanaraman: Washington University School of Medicine
Monique R. Heitmeier: Washington University School of Medicine
Vijay K. Samineni: Washington University School of Medicine

Nature Communications, 2025, vol. 16, issue 1, 1-18

Abstract: Abstract 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.

Date: 2025
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DOI: 10.1038/s41467-025-60360-w

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