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Literature DB >> 30067943

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

Chiso Nwokafor¹, Robert H Singer², Hyungsik Lim³.

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.

Entities: CellLine Chemical Disease Gene Species

Keywords: Intravital microscopy; Transcriptional dynamics; Viral transfer; mRNA transport

Mesh：

Substances：
RNA, Messenger

Year: 2018 PMID： 30067943 PMCID： PMC6351228 DOI： 10.1016/j.ymeth.2018.07.009

Source DB: PubMed Journal: Methods ISSN： 1046-2023 Impact factor: 3.608

29 in total

1. Control of stochasticity in eukaryotic gene expression.

Authors: Jonathan M Raser; Erin K O'Shea
Journal: Science Date: 2004-05-27 Impact factor: 47.728

2. Production and purification of lentiviral vectors.

Authors: Gustavo Tiscornia; Oded Singer; Inder M Verma
Journal: Nat Protoc Date: 2006 Impact factor: 13.491

3. Stereotaxic gene delivery in the rodent brain.

Authors: Ali Cetin; Shoji Komai; Marina Eliava; Peter H Seeburg; Pavel Osten
Journal: Nat Protoc Date: 2006 Impact factor: 13.491

4. An RNA-sequencing transcriptome and splicing database of glia, neurons, and vascular cells of the cerebral cortex.

Authors: Ye Zhang; Kenian Chen; Steven A Sloan; Mariko L Bennett; Anja R Scholze; Sean O'Keeffe; Hemali P Phatnani; Paolo Guarnieri; Christine Caneda; Nadine Ruderisch; Shuyun Deng; Shane A Liddelow; Chaolin Zhang; Richard Daneman; Tom Maniatis; Ben A Barres; Jian Qian Wu
Journal: J Neurosci Date: 2014-09-03 Impact factor: 6.167

5. Human synapsin 1 gene promoter confers highly neuron-specific long-term transgene expression from an adenoviral vector in the adult rat brain depending on the transduced area.

Authors: S Kügler; E Kilic; M Bähr
Journal: Gene Ther Date: 2003-02 Impact factor: 5.250

6. Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window.

Authors: Anthony Holtmaat; Tobias Bonhoeffer; David K Chow; Jyoti Chuckowree; Vincenzo De Paola; Sonja B Hofer; Mark Hübener; Tara Keck; Graham Knott; Wei-Chung A Lee; Ricardo Mostany; Tom D Mrsic-Flogel; Elly Nedivi; Carlos Portera-Cailliau; Karel Svoboda; Joshua T Trachtenberg; Linda Wilbrecht
Journal: Nat Protoc Date: 2009-07-16 Impact factor: 13.491

7. A transcriptome database for astrocytes, neurons, and oligodendrocytes: a new resource for understanding brain development and function.

Authors: John D Cahoy; Ben Emery; Amit Kaushal; Lynette C Foo; Jennifer L Zamanian; Karen S Christopherson; Yi Xing; Jane L Lubischer; Paul A Krieg; Sergey A Krupenko; Wesley J Thompson; Ben A Barres
Journal: J Neurosci Date: 2008-01-02 Impact factor: 6.167

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

1. Control of stochasticity in eukaryotic gene expression.

2. Production and purification of lentiviral vectors.

3. Stereotaxic gene delivery in the rodent brain.

4. An RNA-sequencing transcriptome and splicing database of glia, neurons, and vascular cells of the cerebral cortex.

5. Human synapsin 1 gene promoter confers highly neuron-specific long-term transgene expression from an adenoviral vector in the adult rat brain depending on the transduced area.

6. Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window.

7. A transcriptome database for astrocytes, neurons, and oligodendrocytes: a new resource for understanding brain development and function.

8. High-resolution analysis of parent-of-origin allelic expression in the mouse brain.

9. A transgenic mouse for in vivo detection of endogenous labeled mRNA.

10. Whole transcriptome RNA-Seq allelic expression in human brain.

1. High-pulse-energy multiphoton imaging of neurons and oligodendrocytes in deep murine brain with a fiber laser.