It would be interesting to attempt the R751 knockouts using the TargeTron system from Sigma-Aldrich. Two of the three proposed knockouts could be done with this system. Successful knockouts would answer key questions about the predicted functionality of our system.
Self Destructive Plasmid
The most obvious improvement for future work on a self destructive plasmid would be to replace the LVA-tagged I-SceI homing endonuclease in Status: 500 Content-type: text/html
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by a wildtype sequence. I-SceI protein with the wildtype sequence is commercially available and has been shown to work. The restriction site and reporter gene have been shown to work, so these would not require immediate attention. Possibly the restriction site could be integrated into the reporter gene sequence, so that cleavage would immediately block transcription of the reporter gene.
Synthetic Transcriptional Cascade
It is interesting to test the time-delay device including a constitutive LacI generator and incorporate control as planned, in order to have more reliable results. The complete sequences of the biobricks are being confirmed now.
Biosynthetic AND gate
It might be interesting to construct the planned biobricks here in order to corroborate that a time-delay behavior can be achieved using post-transcriptional mechanisms. Moreover, it is promising to obtain a time-delay using both transcriptional and post-transcriptional regulation in which the researchers can have more parameters to tune the time of the delay.
Lock and Key Library
In order to completely characterize the new riboregulators, the constructed plasmids should be enhanced by the addition of a LacI generator and they should be tested. Although the algorithm and the online generator seem to work, future work on the interface with the user should be improved.
The addition of a conjugation delay time would greatly improve the capabilities of our conjugation model. Another interesting feature would be the ability to confine the growth within predefined geometrical structures. This would allow it to simulate the creation of complex spatiotemporal patters.
First of all, we recommend that in the future, scientists and media work closely together to overcome these issues.
Secondly, both for biosafety and for biosecurity the scientific community has expressed its perception towards the risks involved in synthetic biology and they believe that additional regulation can limit or even reduce these risks.
At last, with our survey we aimed to raise the awareness on ethical issues mainly focusing on the extreme application of the reductionist and bottom-up approach. We got great feedback from many participants and when looking at the amount and the content of the open-answers, most people find this subject very interesting and seemed to appreciate the survey. Although no consensus will be formed on questions like whether life can be explained materialistically, the ongoing research in synthetic biology and specifically on the principles of life makes us obliged to keep the debate going and possibly prepare ourselves for whatever science will bring us next.