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

Chiso Nwokafor, Robert H. Singer, Hyungsik Lim

Research output: Contribution to journalArticle

1 Citation (Scopus)

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)
JournalMethods
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Brain
Capsid Proteins
Imaging techniques
Messenger RNA
Photons
Protein Binding
Cerebral Cortex
Actins
Microscopy
Microscopic examination
Cytoplasm
Molecules
Proteins

Keywords

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

ASJC Scopus subject areas

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

Cite this

Imaging cell-type-specific dynamics of mRNAs in living mouse brain. / Nwokafor, Chiso; Singer, Robert H.; Lim, Hyungsik.

In: Methods, 01.01.2018.

Research output: Contribution to journalArticle

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