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From 2010.igem.org

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Project Overview

Speedy reporter

Speedy switch

Speedy protein degrader

Experiments and Parts

Applications

F.A.Q

About Us

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Contents 1 Project overview by animation 2 Motivation 3 Overview 3.1 Design 4 Acknowledgements

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 just don't fit together in a circuit, and we can't tell without spending time finding out through experimentation.

Even, the current iGEM trend is to create larger and larger circuits, curcuits 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 then 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 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 create limitations in the development of synthetic biology

With this in mind, we are interested in more specific design rules of a genetic circuit. We want to look closely at the central dogma, and more specifically, mRNA.

Modern research is focused on using genes as a reporter, but we want to quantitative description of gene expression in both space and time. Base on the more detail information of gene expression, we can know the interaction between parts in vivo. However, studying the gene expression mechanism by traditional methods takes too much time. To reduce the times of dealing this problem we have come up with our project: SpeedyBac.

Overview

For iGEM2010, the NYMU-Taipei team has created a novel assay that speeds up the expression cycle of a gene. We have designed reporting assays that are faster than conventional methods while revealing the amount and location of mRNA. We have also integrated a faster inducible switch which can switch on/off of protein translation. Finally, we build a speedy degradation system for stop the signal from gene expression specificity and quickly. Combined, these allow us to study mRNA quicker, and better, while reducing the interference of protein translation.

Design

To achieve our goals, our design can split into three parts:

• Speedy switch
  • Faster production of protein by inducing the translation of pre-transcribed RNA molecules.
• Speedy reporter
  • Using mRNA aptamers and split GFP-eIF4A reporter systems to show promoter activity faster.
• Speedy protein degrader
  • Fast, specific, and constitutive proteolysis achieved by engineering fluorescent proteins with LVA tags.

Acknowledgements

Dr. Chris Proud ,for providing us eIF4A for experiment

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