Sequence-Independent RNA Sensing in Living Mammalian Cells

Area of Science:

• Molecular Biology
• RNA Biology
• Biotechnology

Background:

• Adenosine deaminases acting on RNA (ADARs) are utilized in cellular sensors to link target RNA presence to payload translation.
• Existing ADAR-based sensors require specific sequence motifs for stop-codon editing, limiting their application.
• Sensing short RNA sequences, like viral RNAs or splice isoforms, is challenging with current methods.

Purpose of the Study:

• To address the limitations of existing ADAR-based RNA sensors.
• To develop a modular RNA sensor system for detecting short RNA subsequences.
• To enable programmable RNA detection in living cells.

Main Methods:

• Designed modular RNA sensors (modulADAR) with adaptable regions for target transcript hybridization and stem-loop structures for ADAR editing.
• Optimized the modulADAR system for enhanced performance.
• Applied the optimized modulADAR to detect short RNA subsequences.

Main Results:

• Successfully developed and optimized the modulADAR system.
• Demonstrated the ability of modulADAR to detect short RNA subsequences.
• Overcame the motif-dependency of previous ADAR-based sensors.

Conclusions:

• ModulADAR offers a versatile platform for programmable RNA detection in living cells.
• This technology expands the scope of RNA sensing applications, including viral RNA detection and splice isoform differentiation.
• ModulADAR is anticipated to advance basic research and therapeutic development.