Split aminoacyl-tRNA synthetases for proximity-induced stop codon suppression

Area of Science:

• Synthetic Biology
• Molecular Biology
• Biotechnology

Background:

• Gene expression regulation is crucial for biotechnology and therapeutics.
• Existing tools primarily target transcriptional control, with limited options for translational regulation.

Purpose of the Study:

• To engineer novel synthetic biology tools for controlling gene translation.
• To develop split orthogonal aminoacyl-tRNA synthetases (o-aaRS) for precise gene expression modulation.

Main Methods:

• Designed and engineered split o-aaRS using chemically induced dimerization domains.
• Utilized small molecules (rapamycin, abscisic acid) to control o-aaRS activity.
• Demonstrated function in *Escherichia coli* and human cells.

Main Results:

• Split o-aaRS conditionally suppress stop codons in response to specific molecular inputs.
• These systems function as genetically encoded AND gates, requiring dual molecular signals for activation.
• Validated split o-aaRS as biosensors for detecting protein-protein interactions relevant to cancer and viral infections (e.g., SARS-CoV-2).

Conclusions:

• Split o-aaRS provide a novel platform for precise translational control of gene expression.
• The developed tools offer versatile applications in synthetic biology, biosensing, and therapeutic development.