Visualizing RNA dynamics in live cells with bright and stable fluorescent RNAs

Area of Science:

• Molecular Biology
• Biochemistry
• Cell Biology

Background:

• Fluorescent RNAs (FRs) are crucial for visualizing cellular RNA but face limitations in brightness and spectral diversity.
• Existing FRs struggle with applications in live mammalian cells and in vivo due to low signal and limited dye compatibility.

Discussion:

• Peppers offer monomeric, bright, and stable fluorescent RNA aptamers with emission spanning cyan to red.
• These FRs enable robust imaging of diverse RNA species in live cells with minimal impact on RNA processes.
• The study quantifies protein and mRNA levels, revealing heterogeneity in single-cell translation kinetics.

Key Insights:

• Development of Peppers, a new class of fluorescent RNA aptamers with broad spectral range and enhanced brightness.
• Demonstration of Peppers for live-cell imaging of RNA transcription, localization, and translation with high fidelity.
• Application of Peppers in advanced imaging techniques, including CRISPR display for genomic loci and super-resolution microscopy.

Outlook:

• Peppers are poised to become valuable tools for live imaging of cellular RNAs, facilitating deeper understanding of RNA biology.
• Potential for Peppers to be adapted for in vivo imaging and diagnostics.
• Future research may explore further expansion of the spectral range and functionalities of these fluorescent RNA aptamers.