Imaging cell-type-specific dynamics of mRNAs in living mouse brain.
Methods
; 157: 100-105, 2019 03 15.
Article
em En
| MEDLINE
| ID: mdl-30067943
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 ß-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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Encéfalo
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RNA Mensageiro
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Hibridização in Situ Fluorescente
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Rastreamento de Células
Limite:
Animals
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article