Difference between revisions of "Team:LambertGA"

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<h2> Characterization of Nonlysosomal Proteolysis <h2>
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<h2> Characterization of Nonlysosomal Proteolysis </h2>
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<h2>Tuning Biosensors in a Switch-like Fashion<h2>
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<p style="font-size: 20px;">Three million children die every year as a result of micronutrient deficiencies. The World Health Organization estimates that over 2 billion people worldwide are at risk and the Center for Disease Control estimates the costs of testing to be in the millions of dollars. Simple to use, inexpensive biosensors are an area of great interest in synthetic biology. Quite simply they could save millions of lives by providing inexpensive options to address micronutrient deficiencies while saving millions of dollars. The focus of this year’s Lambert’s iGEM team was to research the issues within the area of biosensors and engineer cost effective solutions for both scientists engineering the biosensors and the end users. <br>
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<p style="font-size: 20px;">Micronutrient deficiencies are undoubtedly one of the leading causes of death in the world. Today, according to the WHO, more than 2 billion people - over 30% of the world’s population - suffer from micronutrient deficiencies. However, diagnosing these deficiencies has proved to be extremely expensive and in some cases, very time consuming. To address this growing concern, medical professionals and synthetic biologists have created simple, inexpensive “biosensors” that serve as handy diagnostic tests to use in the field. Nevertheless, these versatile tools still suffer many errors that often yield inaccurate results and even lead to faulty diagnosis. With this in mind, the 2016 Lambert iGEM Team focused primarily on how to enhance existing biosensors and subsequently aid in the diagnosis of micronutrient deficiencies throughout the world.
 
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Biosensors often rely on the response of selection promoters to turn on or off protein expression.  The concentration of proteins in a cell is determined by both the amount synthesized and the amount degraded. Thus, protein degradation is a crucial aspect of maintaining intramolecular equilibrium. A class of ATPases known as AAA+ Proteins involves a well-known proteolysis mechanism known as ClpXP in which ClpX unfolds and translocates a tagged protein into a sequestered proteolytic compartment in ClpP. 
 
 
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Accordingly, we studied protein degradation and devised a “Switch” that prevents the overexpression of specific reporters in biosensors in order to fine tune the overall accuracy of biosensors. By using SsrA degradation tags, we characterized ClpXP, a protease involved in non-lysosomal proteolysis in many prokaryotes, to quantify the relative strength of degradation in GFP and purple chromoproteins.
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We engineered an inducible  construct in which ClpXP will degrade a chromoprotein upon induction. Our system will first be tested in well characterized using Green Fluorescent Protein to establish baseline data. Chromo-protein expression data will be gathered using a device that can quantify the color of the light reflected by the chromoprotein before and after induction. This will ultimately allow us to measure the relative strength of degradation and further characterize a well-known proteolysis mechanism. Our characterization of ClpXP will serve as a precursor for controlled protein delivery in a switch for biosensors.
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Revision as of 22:29, 18 October 2016

Characterization of Nonlysosomal Proteolysis

Micronutrient deficiencies are undoubtedly one of the leading causes of death in the world. Today, according to the WHO, more than 2 billion people - over 30% of the world’s population - suffer from micronutrient deficiencies. However, diagnosing these deficiencies has proved to be extremely expensive and in some cases, very time consuming. To address this growing concern, medical professionals and synthetic biologists have created simple, inexpensive “biosensors” that serve as handy diagnostic tests to use in the field. Nevertheless, these versatile tools still suffer many errors that often yield inaccurate results and even lead to faulty diagnosis. With this in mind, the 2016 Lambert iGEM Team focused primarily on how to enhance existing biosensors and subsequently aid in the diagnosis of micronutrient deficiencies throughout the world.

Accordingly, we studied protein degradation and devised a “Switch” that prevents the overexpression of specific reporters in biosensors in order to fine tune the overall accuracy of biosensors. By using SsrA degradation tags, we characterized ClpXP, a protease involved in non-lysosomal proteolysis in many prokaryotes, to quantify the relative strength of degradation in GFP and purple chromoproteins.