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

Chiso Nwokafor; Robert H. Singer; Hyungsik Lim

doi:10.1016/j.ymeth.2018.07.009

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

Chiso Nwokafor, Robert H. Singer, Hyungsik Lim

Cell Biology

Research output: Contribution to journal › Article › peer-review

3 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 language	English (US)
Pages (from-to)	100-105
Number of pages	6
Journal	Methods
Volume	157
DOIs	https://doi.org/10.1016/j.ymeth.2018.07.009
State	Published - Mar 15 2019

Keywords

Intravital microscopy
Transcriptional dynamics
Viral transfer
mRNA transport

ASJC Scopus subject areas

Molecular Biology
General Biochemistry, Genetics and Molecular Biology

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10.1016/j.ymeth.2018.07.009

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title = "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 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.",

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T1 - Imaging cell-type-specific dynamics of mRNAs in living mouse brain

AU - Nwokafor, Chiso

AU - Singer, Robert H.

AU - Lim, Hyungsik

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N2 - 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.

AB - 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.

KW - Intravital microscopy

KW - Transcriptional dynamics

KW - Viral transfer

KW - mRNA transport

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VL - 157

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JO - Methods

JF - Methods

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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