BME494 Project Group4: Difference between revisions
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==BACKGROUND== | ==BACKGROUND== | ||
[[Image:Geneswitch.png|thumb|frame|left|Circuit diagram of a genetic toggle switch. Image source: http://2009.igem.org/Team:Cambridge]] | [[Image:Geneswitch.png|thumb|frame|left|Circuit diagram of a genetic toggle switch. Gene product A represses promoter b and silences B expression. Image source: http://2009.igem.org/Team:Cambridge]] | ||
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Revision as of 12:42, 13 March 2012
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ABSTRACTOur construct is a light-responsive genetic toggle switch. The construct’s design was inspired by the toggle switch introduced during Unit 1. Our version improves upon that design by exchanging light for ligand molecules as a triggering mechanism for the switch between “ON” and “OFF” states. Light is advantageous over substrate-coupling mechanisms in applied systems because of enhanced control and efficiency during signal exposure. Our light-responsive toggle switch is superior to previous optogenetic methods because gene expression is self-propagated, and continues in the absence of the light signal once initiated. Our construct contains two core components: (1) a blue light responsive promoter derived from the plant Arabidopsis thaliana that was modified into BioBrick format, and (2) a green light promoter system previously used by the 2011 Freiburg iGem team. Our construct was designed using the red fluorescent protein reporter gene, which could be replaced by any gene of interest for a plug-and-play genetic engineering application.
BACKGROUNDA toggle switch is an important design motif in synthetic biology as it allows for the use of different constructs dependent on the active chemicals and gene sequences involved. This allows for multiple constructs to be activated at different times dependent on the wants of the synthetic biologist who created the BioBrick. Expression levels of different gene sequences can be controlled by various conditions such as the presence of ligand molecules in culture media. We improved upon the genetic toggle switch by designing a toggle mechanism that is initiated in the presence of two different wavelengths of light (green and blue light). This allows us to control the rate of reaction of these constructs simply by imploring specific wavelengths that are read by a gene sequence sensitive to that wavelength. The control of this construct is given to the individual performing the experiment as they can induce a gene of interest by simply shining a specific wavelength of blue light onto the construct. In the case of this period of the process, the sequence simply glows red when it is being induced, however, this can be changed at any time to induce nearly any gene of interest. When the construct is completed and all data is collected, the sequence can be terminated with the use of a green light which induces specific aspects of the construct to be activated, and eliminating previous processes.
PROOF OF CONCEPT DESIGN
Prev: 5’-GTT TCT TCC TGC AGC GGC CGC TAC TAG TAA TCT CTA TCG 3’
TESTING
HUMAN PRACTICESOUR TEAM
Your area of study/ academic program/ major, why you are taking BME494, and something interesting about yourself. You may add a link to your personal OWW page.
Your area of study/ academic program/ major, why you are taking BME494, and something interesting about yourself. You may add a link to your personal OWW page.
Your area of study/ academic program/ major, why you are taking BME494, and something interesting about yourself. You may add a link to your personal OWW page.
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