Team:HZAU-China/Results
Results
In this project, we achieved that control cell in the aspect of cell replication, proved that the feasibility of using CRISPR to regulate cell cycle(Figure 1.), and further regulate it with light(Figure 2.). A reverse regulation of our system is demonstrated by OD measurement and qPCR (Figure 3.). At the same time, a corresponding software and hardware are developed to achieve the computer controlled cell cycle. The software is used to predict the internal replication procession and control the hardware to achieve reall-time regulation. For more information please view software and hardware
Notes
When using the CcaS-CcaR system please note that the atc concentration is different from the former concentration 200ng/ml. After transformation of two plasmids related to CcaS-CcaR system into bacteria, it becomes more sensitive to aTc. A final concentration of 200ng/ml will be toxic to the bacteria, and a working concentration as 0.1~1ng/ml is recommended.
In a light-control system, the exist of light sensing molecule is very important, but it is also very neglectable.
Future plan
The future plan including two aspects. One is further developing the system. We plan to developing CcaS-CcaR CRISPRi system by knocking it into genome to make it more stable. At the same time, we hope that we can make the pMag-nMag system work because this system is smaller and less effect on bacterial metabolism. The other aspect is using this system for several certain applications, like characterize the stability of genome gene expression after synchronizer, and so on.
Notes
When using the CcaS-CcaR system please note that the atc concentration is different from the former concentration 200ng/ml. After transformation of two plasmids related to CcaS-CcaR system into bacteria, it becomes more sensitive to aTc. A final concentration of 200ng/ml will be toxic to the bacteria, and a working concentration as 0.1~1ng/ml is recommended.
In a light-control system, the exist of light sensing molecule is very important, but it is also very neglectable.
Future plan
The future plan including two aspects. One is further developing the system. We plan to developing CcaS-CcaR CRISPRi system by knocking it into genome to make it more stable. At the same time, we hope that we can make the pMag-nMag system work because this system is smaller and less effect on bacterial metabolism. The other aspect is using this system for several certain applications, like characterize the stability of genome gene expression after synchronizer, and so on.
