Line 27: | Line 27: | ||
<br>It contains two parts.<br><br> | <br>It contains two parts.<br><br> | ||
− | <font size=" | + | <font size="5" color="#C66C63" face="courier"><b>The switch</b></font><br> One is a light-repressed system containing a single light repressor LexA-VVD, and a ColE promoter containing a SOS operator sequence for LexA binding, regulates the expression of target genes. |
− | <br><p style="text-align:center"><img src="https://static.igem.org/mediawiki/2018/b/b9/T--Tongji_China--picture-Safety-Light_Control-1.png" width=" | + | <br><p style="text-align:center"><img src="https://static.igem.org/mediawiki/2018/b/b9/T--Tongji_China--picture-Safety-Light_Control-1.png"width="40%" style="float: left;"></p> Upon light exposure, the light-switchable repressor LEVI is induced to form homodimers, which then bind to the operator sequence and repress the activity of a promoter by blocking the attachment of RNA polymerase to the promoter. Removal of blue light results in gradual dissociation of the dimers and transcription activation. I repressor, which can repress the activity of the R-O12 promoter, is under the control of the LightOff system. Upon light illumination, LEVI homodimerizes, represses the expression of cI repressor and initiates target gene expression. |
<br><br> | <br><br> | ||
− | <font size=" | + | <font size="5" color="#C66C63" face="courier"><b>The Killer</b></font><br> |
<img src="https://static.igem.org/mediawiki/2018/8/85/T--Tongji_China--picture-Safety-Light_Control-2.png" width="30%" style="float: right;"> Another is for the target gene to kill the bacteria, there we use a suicide gene that encodes Supernova, which is a mutant form of well-known KillerRed, the first engineered genetically-encoded photosensitizer. Photosensitizer contains chromophores that generate reactive oxygen species (ROS) upon illumination, and it is commonly used for illumination-induced destruction of targeted cells. | <img src="https://static.igem.org/mediawiki/2018/8/85/T--Tongji_China--picture-Safety-Light_Control-2.png" width="30%" style="float: right;"> Another is for the target gene to kill the bacteria, there we use a suicide gene that encodes Supernova, which is a mutant form of well-known KillerRed, the first engineered genetically-encoded photosensitizer. Photosensitizer contains chromophores that generate reactive oxygen species (ROS) upon illumination, and it is commonly used for illumination-induced destruction of targeted cells. | ||
+ | <br><br> | ||
<p style="text-align:center"><img src="https://static.igem.org/mediawiki/2018/b/b6/T--Tongji_China--picture-Safety-Light_Control-3.png" width="100%"></p> In general, it forms a light switch and a killer to control the lives of our bacteria, it confirms the safety of our whole project. | <p style="text-align:center"><img src="https://static.igem.org/mediawiki/2018/b/b6/T--Tongji_China--picture-Safety-Light_Control-3.png" width="100%"></p> In general, it forms a light switch and a killer to control the lives of our bacteria, it confirms the safety of our whole project. | ||
<br>Actually our bacteria release in the gut environment for several hours, then they can be eliminated by our immune system. Our project uses this method to ensure the bacteria to be dead when releasing in the environment. It can be safe for human and friendly to the environment. | <br>Actually our bacteria release in the gut environment for several hours, then they can be eliminated by our immune system. Our project uses this method to ensure the bacteria to be dead when releasing in the environment. It can be safe for human and friendly to the environment. | ||
+ | <br><br><br> | ||
+ | <font face=courier, size=4>References:<br> | ||
+ | <font size=2, font face=Arial>1.Chen X, Liu R, Ma Z, et al. An extraordinary stringent and sensitive light-switchable gene expression system for bacterial cells. Cell Res. 2016;26:854–857. doi: 10.1038/cr.2016.74.<br> 2.Trewin AJ, Berry BJ, Wei AY, et al. Light-induced oxidant production by fluorescent proteins. Free Radic Biol Med. 2018 Nov 20;128:157-164. doi: 10.1016. | ||
+ | |||
</div> | </div> |
Revision as of 14:07, 11 October 2018
Safety
Light-off system
We use the light-off system from 2017 SSTi-SZGD to kill our bacteria after effecting.
It contains two parts.
The switch
One is a light-repressed system containing a single light repressor LexA-VVD, and a ColE promoter containing a SOS operator sequence for LexA binding, regulates the expression of target genes.
Upon light exposure, the light-switchable repressor LEVI is induced to form homodimers, which then bind to the operator sequence and repress the activity of a promoter by blocking the attachment of RNA polymerase to the promoter. Removal of blue light results in gradual dissociation of the dimers and transcription activation. I repressor, which can repress the activity of the R-O12 promoter, is under the control of the LightOff system. Upon light illumination, LEVI homodimerizes, represses the expression of cI repressor and initiates target gene expression.
The Killer
Another is for the target gene to kill the bacteria, there we use a suicide gene that encodes Supernova, which is a mutant form of well-known KillerRed, the first engineered genetically-encoded photosensitizer. Photosensitizer contains chromophores that generate reactive oxygen species (ROS) upon illumination, and it is commonly used for illumination-induced destruction of targeted cells.
In general, it forms a light switch and a killer to control the lives of our bacteria, it confirms the safety of our whole project.
Actually our bacteria release in the gut environment for several hours, then they can be eliminated by our immune system. Our project uses this method to ensure the bacteria to be dead when releasing in the environment. It can be safe for human and friendly to the environment.
References:
1.Chen X, Liu R, Ma Z, et al. An extraordinary stringent and sensitive light-switchable gene expression system for bacterial cells. Cell Res. 2016;26:854–857. doi: 10.1038/cr.2016.74.
2.Trewin AJ, Berry BJ, Wei AY, et al. Light-induced oxidant production by fluorescent proteins. Free Radic Biol Med. 2018 Nov 20;128:157-164. doi: 10.1016.
It contains two parts.
The switch
One is a light-repressed system containing a single light repressor LexA-VVD, and a ColE promoter containing a SOS operator sequence for LexA binding, regulates the expression of target genes.
Upon light exposure, the light-switchable repressor LEVI is induced to form homodimers, which then bind to the operator sequence and repress the activity of a promoter by blocking the attachment of RNA polymerase to the promoter. Removal of blue light results in gradual dissociation of the dimers and transcription activation. I repressor, which can repress the activity of the R-O12 promoter, is under the control of the LightOff system. Upon light illumination, LEVI homodimerizes, represses the expression of cI repressor and initiates target gene expression.
The Killer
Another is for the target gene to kill the bacteria, there we use a suicide gene that encodes Supernova, which is a mutant form of well-known KillerRed, the first engineered genetically-encoded photosensitizer. Photosensitizer contains chromophores that generate reactive oxygen species (ROS) upon illumination, and it is commonly used for illumination-induced destruction of targeted cells.
In general, it forms a light switch and a killer to control the lives of our bacteria, it confirms the safety of our whole project.
Actually our bacteria release in the gut environment for several hours, then they can be eliminated by our immune system. Our project uses this method to ensure the bacteria to be dead when releasing in the environment. It can be safe for human and friendly to the environment.
References:
1.Chen X, Liu R, Ma Z, et al. An extraordinary stringent and sensitive light-switchable gene expression system for bacterial cells. Cell Res. 2016;26:854–857. doi: 10.1038/cr.2016.74.
2.Trewin AJ, Berry BJ, Wei AY, et al. Light-induced oxidant production by fluorescent proteins. Free Radic Biol Med. 2018 Nov 20;128:157-164. doi: 10.1016.