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Team:Freiburg/Content/Results/Light system
From 2014.igem.org
The light system
One advantage of our system, the spacial resolution by infecting only distinct areas in a mammalian cell culture with our viral vector, was archived by the use of different light systems. We managed to induce the expression of the mCAT-1 receptor in non-murine cell lines by illumination with light of a special wave length. Since, the viral vector only infect cells that express the mCAT-1 receptor on their surfaces, we transferred genes into distinct areas of a otherwise homogenous tissue using this principle.
Function of the light systems
As the function of the light system, we use, is really important for the function of our whole system, we tested two different light systems. The red light system is based on…. whereas the blue light system uses…blubb
Since, it is known that the red light system works better in chinese hamster ovary (CHO) cells, we used these cells to test the function of this light system. In order to investigate the functionality of the blue light system, we tested both, CHO cells and human embryonic kidney (HEK) cells. Two important findings were archived by this comparison: the blue light system works much more efficient than the red light system; and the blue light system works better with HEK cells than with CHO cells.
Test
The blue light system is very effective in activating an existing reporter brought into the cell before illumination. As we know that the duration of illuminating the system with blue light (452 nm) is critical for its efficiency, we tested various time spans for illumination. Our results indicate that five hours of illumination lead to the highes activation level of SEAP expression that was used as a reporter here.
Test
Another important finding was the specificity of the blue light system. As we always have a dark control in our experiments, we managed to have almost no activation of the SEAP reporter in these samples leading to a very low background level in our system.
