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

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 ?-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.

SUBMITTER: Nwokafor C

PROVIDER: S-EPMC6351228 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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

Nwokafor Chiso C Singer Robert H RH Lim Hyungsik H

Methods (San Diego, Calif.) 20180729

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 b ...[more]

PMID: 30067943

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