Team:SJTU-BioX-Shanghai/Composite Part
Composite PartsWe are so glad to present all the composite parts we have built relevant to our multifactorial visualized detection system during this summer.
How our multifactorial visualized detection system work?
A large variety of chromoproteins can be added on the downstream of Target 1 or Target 3. Here we used amilGFP, amilCP, eforRed, cjBlue four chromoproteins in our detection system. To response to change of outer environment, what we need to do is just choosing the right promoter on the upstream of Antisense 1 or 3. In the absence of factors of interest, Antisense won’t be transcribed, thus Target hairpin structure won’t be disrupted and downstream chromoprotein won’t be expressed. In the presence of factors, promoter is activated, leading transcription of Antisense and subsequently Target is disrupted, colors of chromoproteins exhibit.
The two circuits are constructed into the same plasmid pET-Duet1 or pCDF-Duet1 dual-expression vector. Constitutive promoter J23119 and Target are together synthesized as a whole part and the same as Antisense followed by t500. We have confidence that this system can be widely used as alternative to fluorescence proteins for several merits.
Key advantages:
- So far, two environmental factors can be detected by our system via STAR 1 and STAR 3 and the two STAR systems won’t affect each other. Their DNA sequences are completely different and Blast result showed there is no significant similarity.
- Chromoproteins are excellent indicators because they can be seen just by naked eyes, which means no extra equipment, like UV light, is needed.
- We have also built a detection platform including a scaffold to place a smartphone and APP to analyze colors that chromoproteins exhibit according to the concentration of factors.
- More advantages about STAR itself have been discussed by iGEM16_Imperial and for more information about STAR you can refer to here.
- Chappell J, Takahashi MK, Lucks JB. 2015. Creating small transcription activating RNAs. Nat Chem Biol 11:214–220.
- Meyer, S., Chappell, J., Sankar, S., Chew, R., and Lucks, J. B. (2016) Improving fold activation of small transcription activating RNAs (STARs) with rational RNA engineering strategies Biotechnol. Bioeng. 113, 216.
