Imaging cell-type-specific dynamics of mRNAs in living mouse brain

Chiso Nwokafor, Robert H. Singer, Hyungsik Lim

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

We describe a method for visualizing mRNAs in living mouse. Nascent transcripts and cytoplasmic mRNAs were labeled via lentiviral expression of MS2 coat protein (MCP) tagged with fluorescent protein (MCP-XFP) in knock-in mice whose β-actin mRNAs contained MCP binding stem loops (MBS). Then the mRNA molecules were imaged in the live cerebral cortex through an optical cranial window by intravital two-photon microscopy. By means of the controlled expression of MCP-XFP, single mRNA particles could be detected differentially in the nucleus and cytoplasm of a specific cell type. Consequently, this method is useful for investigating the cell-type-dependent dynamics of mRNAs underlying the structure and function of the brain.

Original languageEnglish (US)
Pages (from-to)100-105
Number of pages6
JournalMethods
Volume157
DOIs
StatePublished - Mar 15 2019

Keywords

  • Intravital microscopy
  • Transcriptional dynamics
  • Viral transfer
  • mRNA transport

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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