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DNA Transformer for Visualizing Endogenous RNA Dynamics in Live Cells.

Ying Wan1,2, Ninghao Zhu1, Yi Lu1

  • 1Department of Biomedical Engineering , The Pennsylvania State University , University Park , Pennsylvania 16802 , United States.

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|February 6, 2019
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Researchers developed a DNA probe to track messenger RNA (mRNA) in living cells. This tool reveals how specific mRNA molecules move and are organized, offering new insights into cellular functions.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biophysics

Background:

  • Intracellular RNA functions are complex and regulated by various mechanisms.
  • Visualizing endogenous mRNA dynamics in live cells remains a significant challenge.
  • Understanding mRNA localization and mobility is crucial for gene expression regulation.

Purpose of the Study:

  • To design and validate a novel biosensor for visualizing endogenous mRNA dynamics in live human cells.
  • To investigate the spatial organization and mobility of Delta-like ligand 4 (Dll4) mRNA.
  • To establish a new tool for studying RNA function and regulation in real-time.

Main Methods:

  • Development of a transformable tetrahedral DNA (T-TED) probe with a flexible hinge structure.
  • Integration of Förster resonance energy transfer (FRET) imaging, super-resolution localization, and single particle tracking.
  • Application of the T-TED biosensor in live human pulmonary microvascular endothelial cells.

Main Results:

  • The T-TED probe successfully visualized the abundance, distribution, and mobility of endogenous Dll4 mRNA.
  • Distinct subpopulations of Dll4 mRNA with unique mobility patterns were identified.
  • Dll4 mRNA subpopulations were observed to be spatially organized near endoplasmic reticulum and microtubule networks.

Conclusions:

  • The T-TED biosensor provides an unprecedented capability to monitor endogenous mRNA dynamics in live human cells.
  • This technology offers a valuable tool for advancing the study of RNA biology and regulation.
  • The findings highlight the complex spatial organization and dynamic nature of mRNA within the cellular environment.