"
Team:NYMU-Taipei/Project
From 2010.igem.org
| Home | Project Overview | Speedy reporter | Speedy switch | Speedy protein degrader | Experiments and Parts | Applications | F.A.Q | About Us |
Contents |
Project overview by animation
Motivation
The Biobrick Parts Registry is beginning to overflow with parts. At last count, there are over 14000 parts in the registry. With this many parts, the registry has made it a very easy to design experiments, but for its complexness it has made it very hard to complete experiments. We design experients, order the parts, receive the parts, and ligate them together, only to find failed results. Even if we follow the correct arrangement of parts (i.e. regulator-RBS-coding sequence-terminator), some parts still don't fit together in a circuit; and we can't tell without spending time finding out through experimentation.
Yet, the current iGEM trend is to create larger and larger circuits, circuits that have less and less chance of working together. We've seen iGEM teams get more creative with bacteria, with more ideas that can enhance synthetic biology. But it often ends as ideas and designs. Rather than spending time hoping that the parts we want will fit together, we need to find out how parts interact so that we know which parts work with each other.
Just like we know how IC components work with each other to create a working circuit, we need to find similar design rules in synthetic biology. The lack of these rules creates limitations in the development of synthetic biology. However, another problem arises when we try to find these rules. With current technology, it takes to much to culture cells, much less be able to discover the intricacies of a cell, even one as simple as bacteria
Current research focuses on using genes as reporters to find out the inner workings of transcription control: “What will happen when a promoter is turned on?” In our case, rather than focusing on transcription, we wish to concentrate on gene expression with regards to space and time. With more detail and information on gene expression, we can explore the interaction between parts in vivo.
So with these problems in mind, we created SpeedyBac.
Overview
For iGEM2010, the NYMU-Taipei team has created a novel assay that can speed up the expression cycle of a gene. Furthermore, the assay also reveals the location and quantity of both mRNA and Proteins. Between the mRNA level and protein level of our gene expression cycle, we have integrated a riboswitch that allows us to stop, start and control the translation of protein. Using this switch, we can study mRNA and protein in one cycle without the interference of one on the other. Lastly, we have built a speedy degradation system that stops the gene expression quickly, and cleanly.
Design
To achieve our goals, our design can split into three parts:
- Speedy switch
- Controls and speends up protein translation via a riboswitch between mRNA and protein level of gene expression.
- Speedy reporter
- Using mRNA aptamers and split GFP-eIF4A reporter systems to quickly promoter activity.
- Speedy protein degrader
- Fast, specific, and constitutive proteolysis achieved by engineering fluorescent proteins with LVA tags.
Acknowledgements
Dr. Chris Proud, for providing us pGEX-eIF4A for experiment.
