Transcriptional profiling of mouse projection neurons with VECTORseq

STAR Protoc. 2022 Aug 18;3(3):101625. doi: 10.1016/j.xpro.2022.101625. eCollection 2022 Sep 16.

Abstract

Existing techniques for transcriptional profiling of projection neurons could be applied to only one neuronal population per experiment. To increase throughput, we developed VECTORseq, which repurposes retrogradely infecting viruses to deliver multiplexable RNA barcodes, enabling projection anatomy to be read out in single-cell datasets. In this protocol, we describe the delivery of viral barcodes to mouse brain to label different projection neurons. We then detail single-cell or nuclei isolation for sequencing, followed by the analysis of single-cell sequencing data. For complete details on the use and execution of this protocol, please refer to Cheung et al. (2021).

Keywords: Bioinformatics; Cell isolation; Gene expression; Genomics; Neuroscience; RNAseq; Sequence analysis; Sequencing; Single cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • High-Throughput Nucleotide Sequencing*
  • Interneurons
  • Mice
  • Neurons
  • RNA*
  • Sequence Analysis, RNA

Substances

  • RNA