These technologies are strategies to conceal synthetic gene circuit topologies from unwanted determination by outside parties, with widespread applications in biotechnology, metabolic engineering, medicine, and biomanufacturing.
Synthetic gene circuits have broad applications in biomedical engineering and industrial biotechnology. Given the difficulty in successfully engineering synthetic gene circuits, as well as their strong commercial potential, it is critical to protect against unwanted determination of gene circuit structure and function by outside parties. For example, elucidation of gene circuit design can be easily achieved through DNA sequencing. However, research into protection of gene circuit design is very limited. With the rising use and implementation of synthetic gene circuits, there is a strong need for efficient and effective methods to protect against unauthorized determination of circuit design and function.
The inventors have developed two strategies for concealing the design of synthetic gene circuits. The first method, called “circuit scrambling,” utilizes recombinases to encrypt the topology of genetic circuits. Specifically, the circuit is strategically designed with multiple, overlapping pairs of recombination-recognition sites, corresponding to different recombinases, such that the circuit topology is scrambled until a set of recombinases is applied in a specific order, leading to the correct and functional circuit. This encryption strategy is particularly useful when the circuit is being stored or transferred between parties. In the second approach, called “circuit camouflage”, the circuit topology is obscured by embedding the functional circuit with dummy components. Introduction of a molecular key, such as utilization of CRISPR to repress the dummy components, leads to revealing of the true circuit. Furthermore, the circuit design can be “rehidden” upon removal of the molecular key. The inventors have demonstrated successful use of both “circuit scrambling” and “circuit camouflage” to conceal multiple types of gene circuits. As the commercial value of synthetic gene circuits increases, strategies to hide genetic circuit design, including those covered by this invention, will be useful and critical.
- Strategies to prevent unwanted determination of synthetic gene circuit design by outside parties
- Brute-force cracking of recombinase-scrambled or camouflaged circuits is highly inefficient, ensuring effective protection of circuit topology