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JoVE Journal
Genetics
Eine optogenetische Methode zu kontrollieren und analysieren gen Expressionsmuster in Zell-Zell-I...
Eine optogenetische Methode zu kontrollieren und analysieren gen Expressionsmuster in Zell-Zell-I...
JoVE Journal
Genetics
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JoVE Journal Genetics
An Optogenetic Method to Control and Analyze Gene Expression Patterns in Cell-to-cell Interactions

Eine optogenetische Methode zu kontrollieren und analysieren gen Expressionsmuster in Zell-Zell-Interaktionen

Full Text
8,225 Views
07:59 min
March 22, 2018

DOI: 10.3791/57149-v

Akihiro Isomura1,2, Ryoichiro Kageyama1,3,4,5

1Institute for Frontier Life and Medical Sciences,Kyoto University, 2Japan Science and Technology Agency,PRESTO, 3Institute for Integrated Cell-Material Sciences,Kyoto University, 4Graduate School of Medicine,Kyoto University, 5Graduate School of Biostudies,Kyoto University

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article presents a protocol for analyzing cell-to-cell transfer of oscillatory information using optogenetic control and live monitoring of gene expression. The method allows for precise control and observation of gene expression dynamics, providing insights into cellular communication.

Key Study Components

Area of Science

  • Neuroscience
  • Cell Biology
  • Optogenetics

Background

  • Understanding cell communication is crucial for deciphering multicellular systems.
  • The Notch signaling pathway plays a significant role in cellular interactions.
  • Oscillatory information transfer is a key aspect of cellular dynamics.
  • Optogenetic tools enable precise manipulation of gene expression.

Purpose of Study

  • To observe how cells communicate through oscillatory signals.
  • To investigate the dynamics of gene expression in a multicellular context.
  • To develop a platform for testing the functional significance of gene expression programs.

Methods Used

  • Transfection of plasmid vectors into C2C12 cells.
  • Use of Tol2-based optogenetic modules.
  • Live monitoring of gene expression dynamics.
  • Cell counting and plating techniques for transfection.

Main Results

  • Successful transfection of optogenetic modules into target cells.
  • Demonstration of oscillatory information transfer between cells.
  • High precision in controlling gene expression dynamics.
  • Insights into the role of Notch signaling in cellular communication.

Conclusions

  • This protocol provides a valuable tool for studying cell communication.
  • Optogenetic control enhances our understanding of gene expression dynamics.
  • The findings may have implications for understanding multicellular systems.

Frequently Asked Questions

What is the main goal of this study?
The main goal is to observe cell-to-cell transfer of oscillatory information.
How does optogenetics contribute to this research?
Optogenetics allows for precise control and monitoring of gene expression dynamics.
What cell type is used in the experiments?
C2C12 cells are used for transfection and observation.
What is the significance of the Notch signaling pathway?
The Notch signaling pathway is crucial for understanding how cells communicate with each other.
Who developed this new technology?
Akihiro Isomura, a research fellow, developed the technology used in this study.
What are the advantages of this method?
The method provides high precision in controlling and monitoring gene expression dynamics.

Hier präsentieren wir Ihnen ein Protokoll zu analysieren von Zelle zu Zelle Übertragung der oszillierenden Informationen durch optogenetische Kontrolle und Überwachung der Genexpression zu leben. Dieser Ansatz bietet eine einzigartige Plattform, um eine funktionale Bedeutung der dynamischen Ausdruck Genprogramme in mehrzelligen Systeme zu testen.

Das übergeordnete Ziel dieses Experiments ist es, den Zell-zu-Zell-Transfer von oszillatorischer Information durch optogenetische Kontrolle und Live-Monitoring der Genexpression zu beobachten. Diese Methode kann dazu beitragen, wichtige Fragen zu beantworten, wie Zellen über den Notch-Signalweg miteinander kommunizieren, insbesondere wie Zellen die oszillatorischen Informationen untereinander übertragen. Der Hauptvorteil dieser Technik besteht darin, dass wir die Dynamik der Genexpression mit sehr hoher Präzision steuern und überwachen können.

Akihiro Isomura, ein Forschungsstipendiat aus meinem Labor, der diese neue Technologie entwickelt hat, wird das Verfahren vorführen. Um Plasmidvektoren von Tol2-basierten optogenetischen Modulen zusammen mit dem Transposase-Expressionsvektor in C2C12-Zellen zu transfizieren, werden zunächst trypsinisierte Zellen mit einem Zellzähler gezählt. Platte 50.000 C2C12-Zellen pro Well in einer 12-Well-Platte einen Tag vor der Transfektion.

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