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In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

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Published on: March 29, 2019

Ribozyme-Enabled Tissue Specificity (RETS): A System for Precise Gene Expression without Specialized Promoters.

Max M Combest¹, Josh Conlin¹, Vivia Van De Mark²

¹Department of Biology, Colorado State University, Fort Collins, Colorado 80523, United States.

ACS Synthetic Biology

|December 19, 2025

Summary

A new method called Ribozyme-Enabled Tissue Specificity (RETS) allows precise control of gene expression in plants. This technique enables tissue-specific transgene expression without needing known promoters, aiding biological studies and crop engineering.

Keywords:

expression biosensor plant development plant synthetic biology ribozymes tissue-specific expression

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Area of Science:

Plant Biology
Molecular Biology
Synthetic Biology

Background:

Achieving precise tissue-specific gene expression in plants is crucial for biological research and engineering but is often hindered by cryptic enhancers and long prototyping timelines.
Existing methods for controlling transgene expression can be challenging due to promoter identification difficulties and the time required for transgenic line development.

Purpose of the Study:

To develop a novel strategy, Ribozyme-Enabled Tissue Specificity (RETS), for achieving predictable, tissue-specific transgene expression in plants without relying on characterized promoters.
To demonstrate the flexibility, efficiency, and applicability of RETS for various plant science applications.

Main Methods:

Developed RETS utilizing a split self-splicing ribozyme (group I intron from *Tetrahymena thermophila*) for conditional mRNA reconstitution.
Designed RETS to leverage transcriptomic data for targeting specific endogenous mRNAs, enabling conditional transgene expression.
Optimized RETS design for transgene/target swapping, enhanced expression, and evasion of RNA interference.

Main Results:

Successfully demonstrated tissue-specific and dose-dependent transgene expression in *Arabidopsis thaliana* using the RETS system.
Showcased RETS utility in creating genetically encoded biosensors for *in planta* gene expression spatiotemporal pattern analysis.
Validated RETS for engineering tissue-specific alterations in organ size, demonstrating its potential for precise phenotype modification.

Conclusions:

RETS offers a groundbreaking approach for studying native gene expression patterns non-destructively, overcoming limitations of current methods.
The spatiotemporal control of transgene expression via RETS facilitates precision engineering of plant phenotypes, enabling crop enhancement without constitutive expression drawbacks.