Periodic Reporting for period 4 - CyberGenetics (Cybergenetics: Theory and Design Tools for Biomolecular Control Systems)
Reporting period: 2022-02-01 to 2022-12-31
The theory and methods developed in this project thus enable the systematic, rational, and effective feedback control of living cells at the gene level, and lay the foundation for a new field which we call “Cybergenetics”. They also open new research directions in the areas of control theory and estimation. We have on demonstrated our methods on several cybergenetic control systems, each addressing an important application in biotechnology or therapeutics. In one application, the controller uses light to precisely regulate gene expression in bioreactor to enhance bioproduction. In a second application, multiple feedback controllers effectively regulate in parallel a large number of single cells using light. Perhaps the most significant applications of our results involve genetic engineering into living cells robust dynamic molecular control systems that implement integral control to precisely and robustly regulate cell behavior. One of our envisioned application of these advanced controller technologies is the real-time monitoring of dysregulated physiological variables and the corresponding expression of biological effectors to bring these variables back to their normal levels, with important implications for the field of cell therapy.
We have also taken a parallel approach for controlling living cells—this time using a digital computer to carry out the controlling. The computer measures the behavior of living cells in real time, and then based on a sophisticated control algorithm commands a light emitting diode to shine light with a carefully chosen intensity on the cells of interest, which have been especially engineered to respond to this light. By continuously measuring the cellular response and correcting it with light, the computer is able to control precisely the dynamic behavior of these cells. We have advanced this type of control in two ways. In the first, we have instrumented bioreactors so we can control cell populations with light and computer feedback in an effort to make bioproduction much more efficient. In the second approach, we have engineered devices that allow us to shine light on single cells. The resulting platform, called Cyberloop, allows single cells to be controlled independently under the microscope. This has many applications in stem cell differentiation, tissue engineering, and basic biology. As we have developed these platforms, we have also developed the necessary theoretical foundations that allow us to understand how to best use them to achieve robust regulation with good performance.
The work carried out in this project has been widely disseminated through journal articles, conference and workshop presentations, and numerous lectures. We have also filed for two patents to protect the IP developed.