Biomod/2011/TeamJapan/Tokyo/Project/Light irradiated gathering mode

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The light-irradiated gathering mode

In this mode, we utilized UV-switching DNA, which contains loop strusture and azobenzenes. Without UV-irradiated, this DNA device doesn’t trap DNA ciliate's legs but with UV irradiated, azobenzenes are isomerized and this DNA device traps DNA ciliate's legs. With this reaction, DNA ciliates gather at UV-irradiated area where UV-switching DNA attached. By using this system, we think DNA’s movement can be controlled.

Method

To achieve this mode, we observed trapping activity of DNA by using complimentary DNA for deoxyribozyme DNA and checked the switching of UV-switching DNA.
Firstly, we tried to confirm DNA ciliates gathered at the area where the complementary DNAs were attached. We prepared glass plate which DNAs were attached at the upper right in Figure 1. In this glass plate, we spotted 200 um DNA ciliates diffused 1x SSC.
Secondly, we tried to confirm this gathering can be happened on the DNA track. We arrayed complementaly strands of deoxyribozyme of DNA ciliate on the glass plate as complementary strands track.
Thirdly, we confirmed UV-switching DNA device. This DNA device is our original designed. This DNA device is switched by UV and traps DNA ciliates' leg. We checked this DNA device’s switching by electrophoresis. This electrophoresis result is here.
(green spot lights are DNA ciliates’ fluorescent polysterene beads)

Result

Figure1:The confirmation of DNA ciliate gathering at one spot.
Figure1:The confirmation of DNA ciliate gathering at one spot.
Figure2:The result of DNA ciliate gathering at one spot. This figure is taken by using fluorescent phase contrast.
Figure2:The result of DNA ciliate gathering at one spot. This figure is taken by using fluorescent phase contrast.
Figure3:The result of DNA ciliate gathering on DNA track.This figure is taken by using fluorescent phase contrast.
Figure3:The result of DNA ciliate gathering on DNA track.This figure is taken by using fluorescent phase contrast.
In Figure 1, at the upper right of the picture, many DNA ciliates are observed. On the other hand, at the lower left, few DNA ciliates are observed. It means DNA ciliates gathered at the area we attached DNAs.
In Figure 2, most of DNA ciliates were observed on the DNA track. It means DNA ciliates gathered on the DNA track.(Figure1,2)
In conclusion, we succeeded in observing DNA ciliates’ gathering at the area we attached complementary DNA strands for deoxyribozyme legs of DNA ciliate. UV-switching DNA has the same sequence for this complementary DNA. So this complementaly is equal to the open state UV-switching DNA. UV-switching DNA will be able to work as trapping device for DNA ciliate's legs.
It means we cleared the confirming of elements of light-irradiated gathering mode.
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