Team:UNESP Brazil/Implementation

Proposed Implementation


PROPOSED IMPLEMENTATION

     Once we decided that our project would be about storing data in DNA and reading it, we realized how difficult it was to put this idea into practice. To implement the regulatory process, we chose the versatile CRISPR-Cas9 system programmed to both induce (CRISPRa) and repress (CRISPRi) gene expression. Reading the message is a tricky part. Conventionally, DNA is read through sequencing, a time-consuming and access-restricted method. We wanted to be able to read any message in real-time and to make the reading easy and affordable. Therefore, we chose the fluorescent RNA aptamers broccoli and corn to sign a readable output in a binary code system. The RNA aptamers bind to a specific fluorophore generating the fluorescent outputs (green and orange) that can be read by simple and portable fluorometer devices - such as our hardware - and translated as 1 and 0 in the binary code. RNA was chosen as output because a transcription-only process is much faster than the transcription-translation process needed for proteins. Moreover, RNA maturation is almost instantaneous while proteins such as GFP may need up to 30 min to fold and form the fluorophore. Finally, RNAs are recycled faster than proteins in the cell, allowing for a fast process of changing from one code to the next. Altogether, our project can revolutionize the way we store and read information, making it simple and portable.

     Design finished, our team looked for more everyday applications for our project. We then realized that it would be possible to use it to control gene expression temporally without using different inducers for each step. Once the process is triggered, the first event induces the next, and so on. That may be used to engineer complex cell behavior triggered by only one input. For example, to engineer smart probiotics to detect a disease biomarker, report the diagnosis, and produce multiple therapeutics in a particular order. So far, autonomous sequential events have been engineered using cyclic regulation only, mimicking a circadian cycle. However, cyclic regulation only alternates 1s and 0s in the same order continuously. We propose a device that can be programmed to autonomously display the outputs in any desired order, not cyclic. The same concept could be used to design smart biological systems for other applications, such as bioremediation, biofertilization, biopesticides. Additionally, a simplified version of the device could be used for diagnosis from body samples. In that case, a specific biomarker would trigger or not the device resulting in a positive or negative readable output for the pathology.

     More information is also available on our Human Practices page.


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