Transcriptional profiling of mouse projection neurons with VECTORseq

Area of Science:

• Neuroscience
• Molecular Biology
• Genomics

Background:

• Current methods for studying projection neurons are limited to analyzing one population per experiment, hindering large-scale anatomical and transcriptomic studies.
• Efficiently profiling diverse neuronal populations is crucial for understanding complex brain circuitry and function.

Purpose of the Study:

• To develop a novel method, VECTORseq, for multiplexed transcriptional profiling of projection neurons.
• To enable simultaneous analysis of multiple neuronal populations and their projection targets.

Main Methods:

• VECTORseq utilizes retrogradely infecting viruses to deliver unique RNA barcodes to distinct projection neuron populations in the mouse brain.
• The protocol involves viral barcode delivery, single-cell or single-nuclei isolation, RNA sequencing, and subsequent bioinformatic analysis.
• This approach allows for the simultaneous readout of projection anatomy and gene expression profiles.

Main Results:

• VECTORseq successfully labels and distinguishes multiple projection neuron populations based on their viral barcode sequences.
• The method integrates projection targeting information with single-cell RNA sequencing data.
• Enables comprehensive analysis of neuronal connectivity and transcriptomic diversity.

Conclusions:

• VECTORseq significantly increases throughput for studying projection neuron populations.
• This technique provides a powerful tool for dissecting brain circuitry and understanding neuronal diversity.
• Facilitates large-scale mapping of neuronal connections and gene expression patterns.