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RNA Interference

RNA Interference

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Related Experiment Video

Updated: May 28, 2025

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Published on: August 6, 2014

A molecular proximity sensor based on an engineered, dual-component guide RNA.

Junhong Choi^1,2, Wei Chen^1,3, Hanna Liao^1,4

¹Department of Genome Sciences, University of Washington, Seattle, United States.

|February 12, 2025

Summary

We developed P3 editing, a new method linking protein proximity to CRISPR-Cas9 genome editing. This allows precise control over gene editing in synthetic biology circuits within living cells.

Keywords:

CRISPR-Cas genetics genome editing genomics human molecular recording protein-protein interaction synthetic biology

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Area of Science:

Synthetic Biology
Molecular Biology
Genome Engineering

Background:

Synthetic biology aims to create programmable molecular circuits in cells.
Genome editing tools like CRISPR-Cas9 are crucial for these circuits.
Controlling genome editing with molecular events like protein interactions is challenging.

Purpose of the Study:

To develop a novel strategy for programming genome editing using protein-protein proximity.
To enhance the controllability of CRISPR-Cas9 based genome editing for synthetic biology applications.

Main Methods:

Developed 'P3 editing' by engineering the CRISPR-Cas9 dual-component guide RNA.
Linked protein-protein proximity to guide RNA formation.
Activated prime editing and base editing using known and chemically induced protein interactions in human cells.

Main Results:

Demonstrated P3 editing activates CRISPR-Cas9 based prime and base editing.
Showcased control using various protein-protein interactions and chemically induced dimerization.
Explored integration with RNA sensors for RNA-inducible genome editing.

Conclusions:

P3 editing enhances the programmability and controllability of genome editing.
This strategy facilitates the development of sophisticated synthetic molecular circuits.
Enables precise, condition-specific genome modifications within living cells.