Single molecule-sensitive probes for imaging RNA in live cells.

Area of Science:

• Molecular Biology
• Cell Biology
• Biophysics

Background:

• Visualizing native RNA in live cells at the single-molecule level is challenging.
• Existing methods often require genetic modification or lack sensitivity.

Purpose of the Study:

• To develop a novel imaging probe for high-sensitivity visualization of native RNA in living cells.
• To demonstrate the utility of this probe for studying RNA dynamics and interactions.

Main Methods:

• Development of multiply labeled tetravalent RNA imaging probes (MTRIPs).
• Delivery of MTRIPs into live human epithelial cancer cells and primary chicken fibroblasts using streptolysin O.
• Single-molecule imaging of native mRNAs and a non-engineered viral RNA.

Main Results:

• MTRIPs enabled accurate imaging of native mRNAs and a non-engineered viral RNA in live cells.
• The probes facilitated visualization of RNA co-localization with known RNA-binding proteins.
• MTRIPs allowed the study of RNA dynamics and interactions with cellular structures like stress granules.

Conclusions:

• MTRIPs provide a powerful tool for single-molecule visualization of native RNA in live cells.
• This technology opens new avenues for studying RNA biology, including dynamics and interactions.
• The method is applicable to both endogenous and viral RNA in various cell types.