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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Using Genetically Encoded Fluorescent Biosensors for Quantitative In Vivo Imaging.

Akira Yoshinari1, Jacob Moe-Lange2, Thomas J Kleist3

  • 1Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Nagoya, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|November 11, 2020
PubMed
Summary

Fluorescent protein biosensors enable visualization of cellular processes in plants. This study demonstrates using a calcium indicator to track intracellular Ca2+ dynamics in Arabidopsis thaliana.

Keywords:
Arabidopsis root perfusion chamberFRETFluorescence microscopyGenetically encoded fluorescent biosensorscpFP

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Area of Science:

  • Plant biology
  • Molecular imaging
  • Biotechnology

Background:

  • Fluorescent biosensors are crucial for real-time monitoring of cellular activities in living organisms.
  • Fluorescent protein (FP)-based biosensors offer high selectivity and minimal invasiveness for biological research.
  • FP-based biosensors have been widely applied in plant science to study dynamic changes in various cellular components and processes.

Purpose of the Study:

  • To summarize the applications of FP-based biosensors in plant research.
  • To present methods for monitoring intracellular Ca2+ dynamics in plants.
  • To highlight the utility of genetically encoded calcium indicators for in planta studies.

Main Methods:

  • Review of existing literature on FP-based biosensors in plant research.
  • Demonstration of monitoring intracellular Ca2+ dynamics in Arabidopsis thaliana.
  • Utilizing a ratiometric genetically encoded Ca2+ indicator, MatryoshCaMP6s.

Main Results:

  • FP-based biosensors are versatile tools for visualizing diverse cellular events in plants.
  • Intracellular Ca2+ dynamics in Arabidopsis thaliana can be effectively monitored using MatryoshCaMP6s.
  • The study provides a practical approach for studying calcium signaling in plants.

Conclusions:

  • FP-based biosensors are invaluable for advancing plant science research.
  • MatryoshCaMP6s serves as a reliable tool for real-time Ca2+ imaging in planta.
  • This work facilitates further investigation into calcium-mediated processes in plants.