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<a class="zhengwen_disblock">On the transcription level, we used CcaS-CcaR system, which is developed from cyanobacteria and is well used in synthetic biology</a><a class="yinzhu" href="#yinwen_jiaozheng">$^{[3]}$</a><a class="zhengwen_disblock">. The CcaS-CcaR system is a two-component system. Under the green light, CcaS protein will be phosphorylated and the CcaR protein accept this phosphate and dimerize into a transcriptor inducing the transcription of gRNA, in which leading to the inhibition of replication. In the red light, the gRNA will stop transcription and degrade in a short time, and the inhibition will decrease in a short time freeing cell from blocking(Figure 1.). Corresponding results please see HERE.</a> | <a class="zhengwen_disblock">On the transcription level, we used CcaS-CcaR system, which is developed from cyanobacteria and is well used in synthetic biology</a><a class="yinzhu" href="#yinwen_jiaozheng">$^{[3]}$</a><a class="zhengwen_disblock">. The CcaS-CcaR system is a two-component system. Under the green light, CcaS protein will be phosphorylated and the CcaR protein accept this phosphate and dimerize into a transcriptor inducing the transcription of gRNA, in which leading to the inhibition of replication. In the red light, the gRNA will stop transcription and degrade in a short time, and the inhibition will decrease in a short time freeing cell from blocking(Figure 1.). Corresponding results please see HERE.</a> | ||
<a font-size = "10px"><img src="https://static.igem.org/mediawiki/2017/9/91/T--HZAU-China--design_figure2.png" class="tu_1">Figure 2. The CcaS-CcaR system is developed from Synechocystis PCC 6803, and engineered into <i>E.coli</i>. It is a two component system(TCS) in which CcaS can sense green light and autophosphorylate as a membrane-binding protein, and CcaR can be phophorylated by CcaS-P and dimerize into a transcription factor. <br/></a> | <a font-size = "10px"><img src="https://static.igem.org/mediawiki/2017/9/91/T--HZAU-China--design_figure2.png" class="tu_1">Figure 2. The CcaS-CcaR system is developed from Synechocystis PCC 6803, and engineered into <i>E.coli</i>. It is a two component system(TCS) in which CcaS can sense green light and autophosphorylate as a membrane-binding protein, and CcaR can be phophorylated by CcaS-P and dimerize into a transcription factor. <br/></a> | ||
− | <a class="zhengwen_disblock">The protein level of light controlled dCas9 system is based on the split protein and light induced dimerization (LID) protein. By infusing split dCas9 and LID protein together dCas9 can be controlled by light. (Figure 2.) The pMag and nMag developed from fungal is chosen to induce the complement of dCas9<a class="yinzhu" href="#yinwen_jiaozheng">$^{[4]}$</a><a class="zhengwen_disblock">. They are engineered VVD protein using FAD as its light sensing molecule. Under the irritation of blue light, the conformation change of FAD influences the structure of pMag and nMag revealing its dimerization domain. We choose this pair of protein due to its low molecular weight and tunable dynamics. (Figue | + | <a class="zhengwen_disblock">The protein level of light controlled dCas9 system is based on the split protein and light induced dimerization (LID) protein. By infusing split dCas9 and LID protein together dCas9 can be controlled by light. (Figure 2.) The pMag and nMag developed from fungal is chosen to induce the complement of dCas9<a class="yinzhu" href="#yinwen_jiaozheng">$^{[4]}$</a><a class="zhengwen_disblock">. They are engineered VVD protein using FAD as its light sensing molecule. Under the irritation of blue light, the conformation change of FAD influences the structure of pMag and nMag revealing its dimerization domain. We choose this pair of protein due to its low molecular weight and tunable dynamics. (Figue 3. )</a> |
<a><img src="https://static.igem.org/mediawiki/2017/1/18/T--HZAU-China--design_figure3.png" class="tu_1">Figure 3. | <a><img src="https://static.igem.org/mediawiki/2017/1/18/T--HZAU-China--design_figure3.png" class="tu_1">Figure 3. | ||
<a class="zhengwen">These two approaches both can satisfy our need in a certain way, but the former one is simpler but more related to metabolic state of chassis, and the latter one is more difficult to fulfill. We tried two approaches at the same time. For more information please view EXPERIMENT.</a> | <a class="zhengwen">These two approaches both can satisfy our need in a certain way, but the former one is simpler but more related to metabolic state of chassis, and the latter one is more difficult to fulfill. We tried two approaches at the same time. For more information please view EXPERIMENT.</a> |
Revision as of 08:24, 1 November 2017
The protein level of light controlled dCas9 system is based on the split protein and light induced dimerization (LID) protein. By infusing split dCas9 and LID protein together dCas9 can be controlled by light. (Figure 2.) The pMag and nMag developed from fungal is chosen to induce the complement of dCas9$^{[4]}$. They are engineered VVD protein using FAD as its light sensing molecule. Under the irritation of blue light, the conformation change of FAD influences the structure of pMag and nMag revealing its dimerization domain. We choose this pair of protein due to its low molecular weight and tunable dynamics. (Figue 3. ) Figure 3. These two approaches both can satisfy our need in a certain way, but the former one is simpler but more related to metabolic state of chassis, and the latter one is more difficult to fulfill. We tried two approaches at the same time. For more information please view EXPERIMENT.