Target-dependent RNA polymerase as universal platform for gene expression control in response to intracellular...

Area of Science:

• Synthetic Biology
• Molecular Biology
• Genetic Engineering

Background:

• Controlling cellular functions relies on regulating gene expression.
• Existing methods for inducing gene expression are limited by the number of detectable molecules.

Purpose of the Study:

• To develop a novel RNA polymerase system for gene expression control.
• To expand the range of molecules that can induce gene expression.
• To enable advanced genetic circuit construction and cell-specific genome editing.

Main Methods:

• Engineered a Target-dependent RNA polymerase (TdRNAP) that initiates RNA transcription upon binding to a specific intracellular target recognized by an antibody.
• Demonstrated TdRNAP responsiveness to diverse molecules (peptides, proteins, RNA, small molecules) by substituting the fused antibody.
• Constructed orthogonal and multilayer genetic circuits using multiple TdRNAPs.
• Applied TdRNAP for autonomous, cell-specific genome editing triggered by target gene product detection.

Main Results:

• TdRNAPs successfully induced RNA transcription in response to a broad spectrum of intracellular molecules in human cells.
• Demonstrated the ability to create complex, independent genetic circuits with multiple TdRNAPs.
• Achieved precise, cell-specific genome editing autonomously activated by detecting specific cellular targets.
• Showcased the versatility of TdRNAP by responding to various molecular types.

Conclusions:

• Target-dependent RNA polymerase (TdRNAP) significantly expands the molecular repertoire for controlling gene expression.
• TdRNAP offers a versatile genetic toolbox for advanced bioengineering.
• This technology holds promise for future therapeutic applications requiring precise molecular control.