<h2>Suicide system for all the engineered <i>E. coli</i></h2>
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<b>QS (quorum sensing) suicide system</b><br><br>
This system (QS suicide system) is a system that will self-start when our engineering bacteria are lost to the nature. How does this system work? Before explaining the operation of this system, we need to know that there are lots of acyl-homomserine lactones (AHL) as it is QS signaling compounds of Gram-negative bacteria which are frequent colonizers of the nature. Also, it is noteworthy that our engineering bacteria have been artificially added with a special protein, LUXR, before doing research. In this case, whenever the engineering bacteria are lost in to the nature, the LUXR in the engineering bacteria will meet AHL as it is so common in the nature, and they will combine together to form LUXR-AHL complex. This complex will activate the cas9 in the engineering bacteria, which we added artificially. Cas9 is a 160 kilodalton protein which plays a vital role in the immunological defense of certain bacteria against DNA viruses and plasmids, and is heavily utilized in genetic engineering applications. Cas9 we used contains a specific sgRNA which is a specific RNA sequence that recognizes the target DNA region of interest and directs the Cas nuclease there for editing, so that they can function together to cut ATP synthase in the staphylococcus aureus in order to disfunction staphylococcus aureus. In addition, the sequence of sgRNA is AAAGTCGCAATTGTATGCAC. How we find the this specific sgRNA? First, we used uniprot to ascertain the specific ATP, which needs to be destroyed. The entry number of this specific ATP is P68699 in the uniprot, and the name is ATP synthase submit c. Second, we found the complete sequence of this ATP, and then use www.idtdna.com, a sgRNA design tool, to design this specific sgRNA. In this case, as we add this QS suicide system in to engineering bacteria, it can take precaution against the danger of the engineering bacteria’s leakage. The exact processes are shown on the figure below.
This system (QS suicide system) is a system that will self-start when our engineering bacteria are lost to the nature. How does this system work? Before explaining the operation of this system, we need to know that there are lots of acyl-homomserine lactones (AHL) as it is QS signaling compounds of Gram-negative bacteria which are frequent colonizers of the nature. Also, it is noteworthy that our engineering bacteria have been artificially added with a special protein, LUXR, before doing research. In this case, whenever the engineering bacteria are lost in to the nature, the LUXR in the engineering bacteria will meet AHL as it is so common in the nature, and they will combine together to form LUXR-AHL complex. This complex will activate the cas9 in the engineering bacteria, which we added artificially. Cas9 is a 160 kilodalton protein which plays a vital role in the immunological defense of certain bacteria against DNA viruses and plasmids, and is heavily utilized in genetic engineering applications. Cas9 we used contains a specific sgRNA which is a specific RNA sequence that recognizes the target DNA region of interest and directs the Cas nuclease there for editing, so that they can function together to cut ATP synthase in the <i>Staphylococcus aureus</i> in order to disfunction <i>Staphylococcus aureus</i>. In addition, the sequence of sgRNA is AAAGTCGCAATTGTATGCAC. How we find the this specific sgRNA? First, we used uniprot to ascertain the specific ATP, which needs to be destroyed. The entry number of this specific ATP is P68699 in the uniprot, and the name is ATP synthase submit c. Second, we found the complete sequence of this ATP, and then use www.idtdna.com, a sgRNA design tool, to design this specific sgRNA. In this case, as we add this QS suicide system in to engineering bacteria, it can take precaution against the danger of the engineering bacteria’s leakage. The exact processes are shown on the figure below.
