Genetically Encoded Catalytic Hairpin Assembly for Sensitive RNA Imaging in Live Cells

Area of Science:

• Molecular Biology
• Biotechnology
• Nanotechnology

Background:

• DNA and RNA nanotechnology advances dynamic molecular devices.
• Enzyme-free nucleic acid circuits, like catalytic hairpin assembly (CHA), show promise for bioanalysis.
• Intracellular delivery and degradation challenges limit synthetic nucleic acid circuits.

Purpose of the Study:

• To develop a genetically encoded RNA-based CHA circuit for sensitive intracellular RNA imaging.
• To overcome limitations of synthetic nucleic acid circuits for long-term cellular applications.
• To engineer a programmable system for imaging diverse RNA targets.

Main Methods:

• Utilized a genetically encoded RNA-based catalytic hairpin assembly circuit.
• Employed a split version of the Broccoli RNA aptamer as a fluorogenic reporter.
• Engineered the circuit for programming to image various target RNA sequences.

Main Results:

• Achieved sensitive detection of target RNAs inside living cells.
• Demonstrated that one target RNA can catalytically trigger fluorescence from multiple Broccoli aptamers.
• Developed a programmable system for versatile RNA imaging applications.

Conclusions:

• Genetically encoded RNA circuits offer a powerful alternative for long-term cellular analysis and regulation.
• The developed RNA-based CHA circuit enables sensitive and programmable RNA imaging in living cells.
• This design principle facilitates the creation of diverse RNA circuits for cellular applications.