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Cell-type and subcellular compartment-specific APEX2 proximity labeling reveals activity-dependent nuclear proteome dynamics in the striatum

Nat Commun. 2021-08; 
V Dumrongprechachan, R B Salisbury, G Soto, M Kumar, M L MacDonald, Y Kozorovitskiy
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Custom Vector Construction H2B-APEX2-P2A-EGFP were synthesized by Genscript and subcloned into pAAV-EF1a-DIO-WPRE-hGH vector (a gift from Dr. Karl Deisseroth Get A Quote

摘要

The vertebrate brain consists of diverse neuronal types, classified by distinct anatomy and function, along with divergent transcriptomes and proteomes. Defining the cell-type specific neuroproteomes is important for understanding the development and functional organization of neural circuits. This task remains challenging in complex tissue, due to suboptimal protein isolation techniques that often result in loss of cell-type specific information and incomplete capture of subcellular compartments. Here, we develop a genetically targeted proximity labeling approach to identify cell-type specific subcellular proteomes in the mouse brain, confirmed by imaging, electron microscopy, and mass spectrometry. We virally... More

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