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

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 o...

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Main Authors: Lite Yang, Fang Liu, Hannah Hahm, Takao Okuda, Xiaoyue Li, Yufen Zhang, Vani Kalyanaraman, Monique R. Heitmeier, Vijay K. Samineni
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60360-w
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author Lite Yang
Fang Liu
Hannah Hahm
Takao Okuda
Xiaoyue Li
Yufen Zhang
Vani Kalyanaraman
Monique R. Heitmeier
Vijay K. Samineni
author_facet Lite Yang
Fang Liu
Hannah Hahm
Takao Okuda
Xiaoyue Li
Yufen Zhang
Vani Kalyanaraman
Monique R. Heitmeier
Vijay K. Samineni
author_sort Lite Yang
collection DOAJ
description 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.
format Article
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institution DOAJ
issn 2041-1723
language English
publishDate 2025-07-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-208de1f2a1ef417cb413503b2c11c9552025-08-20T03:03:25ZengNature PortfolioNature Communications2041-17232025-07-0116111810.1038/s41467-025-60360-wProjection-TAGs enable multiplex projection tracing and multi-modal profiling of projection neuronsLite Yang0Fang Liu1Hannah Hahm2Takao Okuda3Xiaoyue Li4Yufen Zhang5Vani Kalyanaraman6Monique R. Heitmeier7Vijay K. Samineni8Department of Anesthesiology, Washington University Pain Center, Washington University School of MedicineDepartment of Anesthesiology, Washington University Pain Center, Washington University School of MedicineDepartment of Anesthesiology, Washington University Pain Center, Washington University School of MedicineDepartment of Anesthesiology, Washington University Pain Center, Washington University School of MedicineDepartment of Anesthesiology, Washington University Pain Center, Washington University School of MedicineDepartment of Anesthesiology, Washington University Pain Center, Washington University School of MedicineDepartment of Anesthesiology, Washington University Pain Center, Washington University School of MedicineDepartment of Anesthesiology, Washington University Pain Center, Washington University School of MedicineDepartment of Anesthesiology, Washington University Pain Center, Washington University School of MedicineAbstract 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.https://doi.org/10.1038/s41467-025-60360-w
spellingShingle Lite Yang
Fang Liu
Hannah Hahm
Takao Okuda
Xiaoyue Li
Yufen Zhang
Vani Kalyanaraman
Monique R. Heitmeier
Vijay K. Samineni
Projection-TAGs enable multiplex projection tracing and multi-modal profiling of projection neurons
Nature Communications
title Projection-TAGs enable multiplex projection tracing and multi-modal profiling of projection neurons
title_full Projection-TAGs enable multiplex projection tracing and multi-modal profiling of projection neurons
title_fullStr Projection-TAGs enable multiplex projection tracing and multi-modal profiling of projection neurons
title_full_unstemmed Projection-TAGs enable multiplex projection tracing and multi-modal profiling of projection neurons
title_short Projection-TAGs enable multiplex projection tracing and multi-modal profiling of projection neurons
title_sort projection tags enable multiplex projection tracing and multi modal profiling of projection neurons
url https://doi.org/10.1038/s41467-025-60360-w
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