Inert Pepper aptamer-mediated endogenous mRNA recognition and imaging in living cells.

Area of Science:

• Molecular Biology
• Biotechnology
• Cellular Imaging

Background:

• RNA aptamer/fluorophore systems are crucial for in vivo RNA dynamics studies.
• Existing Pepper-based systems for mRNA imaging require sequence insertion and yield low fluorescence signals.
• These limitations hinder the application of current systems for endogenous mRNA imaging in live cells.

Purpose of the Study:

• To develop an improved RNA aptamer/fluorophore system for sensitive and selective imaging of endogenous mRNAs.
• To overcome the limitations of existing Pepper-based systems, such as sequence insertion requirements and low signal.
• To enable minimal perturbation of target mRNAs during live-cell imaging.

Main Methods:

• Remodeling the original Pepper aptamer to create a tandem array of inert Pepper (iPepper) units.
• Development of a fluorescence turn-on system based on the iPepper array.
• Application of the iPepper system for imaging diverse endogenous mRNA species in live cells.

Main Results:

• The iPepper system demonstrates efficient and selective imaging of various endogenous mRNA types.
• The system achieves high fluorescence signals with minimal agitation of target mRNAs.
• Successful application in live-cell imaging of endogenous mRNA species.

Conclusions:

• The iPepper system represents a significant advancement for endogenous mRNA imaging in live cells.
• It overcomes key limitations of previous aptamer/fluorophore imaging techniques.
• iPepper holds potential as a powerful tool for real-time biological studies and processing in living cells.