A New Platform for Protease-Based Synthetic Biologic Circuit

Synthetic biology and DNA circuits

Synthetic biology has been developed for more than 20 years, scientists have long hoped that synthetic biology could create biomolecular circuits to characterize the state of cells, process the information, and then deliver therapeutic outputs accordingly. The transcription circuit realized by DNA has already achieved precise research results in many aspects. In particular, a DNA circuit that uses ssDNA as the input/output signal through DNA strand displacement provides a useful tool for regulating gene networks and processing molecular information. The newly designed gene is integrated into the cell to express the protein that can recognize the function with the help of various enzymes. A DNA circuit is similar to an electronic circuit, both process input signals through components to control the output of downstream signals. The construction of DNA circuits can simplify and reprogram natural regulatory circuits or introduce artificial laws that do not exist in nature which can provide new solutions for medicine and health, the agricultural environment, and industrial fermentation. But there is a big problem with this circuit: gens must undergo the process of translation before they can form proteins to perform their functions, so there will be problems such as inconvenient delivery and poor operational stability. However, most of the current applications of this discipline are limited to the field of DNA, and few directly synthesize proteins for regulation.

Fig.1 Synthetic gene circuits for theranostic applications.

Protein circuits

As the essential organic matter that constitutes cells, protein is the material basis of life and the main undertaker of life activities. Proteins are involved in every cell and are a vital component of the human body. Therefore, the study of protein circuits has become significant in biologics. Compared with the transcription circuit, the protein circuit can solve the problem of poor stability caused by the translation. However, there are still some other problems in protein circuits. For example, the components of protein circuits are less modular, which makes engineering inconvenient. Therefore, the problem of modularization of the guarantee circuit needs to be solved urgently.

Recently, researchers have made breakthroughs in the study of protein circuits. They have developed a technology platform with an ideal protein circuit that controls protein secretion and cell-cell communication through proteases. The critical regulatory element of this protein circuit is a protease that can cleave any specific sequence. The platform allows the target secreted protein to stay in specific organelles temporarily. The action of proteases disrupts this immobilization, and the target protein is subsequently secreted into the extracellular. This platform is compatible with both protein secretion and the surface display of membrane proteins and could be used to control physiological outputs. Moreover, this platform could sense and process components to respond to internal cell states and external signals via engineered receptors. This platform demonstrates a protein-level control module to directly regulate protein secretion compatible with pre-existing protein components to program therapeutic circuits for cancer immunotherapy and transplantation in the future.

The Advantages of this method include:

• Provide a new approach to cancer immunotherapy
• Regulated almost any extracellular protein
• Protein circuit improves mutation probability in DNA circuits
• Compatible with pre-existing protein-based synthetic circuits

Potential Applications

• Cancer immunotherapy and cell therapies
• Synthetic DNA and protein circuits research
• Cascaded signals research

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Reference

1. Saltepe B, Kehribar ES, Su Yirmibeşoglu SS, Safak Şeker UO. Cellular Biosensors with Engineered Genetic Circuits. ACS Sensors.  2018, 26,3(1):13-26. Doi: 10.1021/acssensors.7b00728.