Provide a faster assay system for exploring the design rules of synthetic biology.
Why do we want to do that?
There are already many genetic parts in the Biobrick Parts Registry and the numbers are growing fast. Every year every igem teams will build genetic circuits based on the parts at partsregistry. But where are the design rules to put these parts into circuits of devices and systems? Apparently, the "Assembly Standards" listed at the partsregistry are only used to connect compatible restriction enzyme cutting sites. They are NOT designing principles. Our iGEM team is very interested in the detailed design rules played in the central dogma; especially those principles for connecting mRNA translation to protein folding. Traditionally, we know about the circuits we made are working or not mostly through the expression of reporter genes. However, it would be much helpful if we could have information of quantitative description of gene expression in both space and time. For these reasons and for the future development of synthetic biology, we just have to speed up the experimental explorations of design rules.
- detect gene expression quantitatively in both space and time.
- specific insight into the flow of genetic information.
- provide speedy ways to report and stop gene expression.
To achieve our specific aim, we have designed a novel reporting device(Speedy reporter) for quickly detectin and measuring the mRNA location and quantity, it can also be used for protein detection. And we design a novel switch (Speedy switch) for control the mRNA translation of gene expression. We have also designed a faster degradation device (Speedy protein degrader); it allows us to regulate the degradation time for studying the mRNAs without the interference from translation and quickly stopping the gene expression.
Our SpeedyBac system is made up of the following three devices:
- Speedy switch
Faster production of protein by inducing the translation of pre-existing mRNA molecules.
- Speedy reporter
Using mRNA aptamers and split GFP-eIF4A reporter designs to detect promoter activity faster.
- Speedy protein degrader
Fast, specific, and constitutive proteolysis achieved by engineering fluorescent proteins tagged with LVA