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<head><meta name="viewport" content="width=device-width, initial-scale=1"><meta http-equiv="Content-Type" content="text/html; charset=UTF-8"><script>function disputeIframe(){setTimeout(function (){self.document.open();self.document.write("<h1><a style='font-weight: normal; color: #000; font-family: arial;' href='" + self.location.href + "' target='_top'>Iframes are not supported on a free plan.</a></h1>");                    self.document.close();},200);} try {if (self!=top){disputeIframe();}}catch(e){disputeIframe();}</script><link href="//d33rxv6e3thba6.cloudfront.net/asset/sites/css/bootstrap3.min.css" rel="stylesheet"><link rel="stylesheet" href="//d33rxv6e3thba6.cloudfront.net/asset/sites/css/font-awesome.min4.css"><link href="//d33rxv6e3thba6.cloudfront.net/asset/sites/css/hover-min.css" rel="stylesheet"><link href="/static/css/style.css?v=2.0.4" rel="stylesheet"><link href="/static/css/blocks.css?v=2.0.4" rel="stylesheet"><link href="/static/css/theme-css/ez.css?v=2.0.4" rel="stylesheet"><link rel="stylesheet" type="text/css" href="//fonts.googleapis.com/css?family=Roboto:100,200,300,400,500,600,700,800,900,100italic,200italic,300italic,400italic,500italic,600italic,700italic,800italic,900italic" /><style  type='text/css' data='Page Custom CSS'></style><script src="//code.jquery.com/jquery-1.12.4.min.js" integrity="sha256-ZosEbRLbNQzLpnKIkEdrPv7lOy9C27hHQ+Xp8a4MxAQ=" crossorigin="anonymous"></script><script> if (!window.jQuery) {  document.write('<script src="//d33rxv6e3thba6.cloudfront.net/asset/sites/js/jquery.js"></scr' + 'ipt>'); }</script><meta name="page-source" site="//sites.simbla.com/52b5a023-1bcf-d512-a412-3351ef830f04" page="description" site-id="593aca5737cad4ed7fa1fc16" page-id="593f0ce46ed040a131fa79eb" master-page-id="" /><link rel="canonical" href="//sites.simbla.com/52b5a023-1bcf-d512-a412-3351ef830f04/description" /></head><body data-spy="scroll" data-target=".navbar-collapse" class="static-page"><div id="windowDiv"><div id="MarginsTop" style="height: 0px;" class="row"></div><div id="bodyContainer" style="width:100%"><div class="simblaEL containerHolder" data-drag="P27" id="P27" style=""><a class="simblaEL anchorElm" data-drag="divCounter" name="Content1" id="Content1" data-id="593f0f2e54ae60a331d6587e"></a><div class="center-bg-image container-fluid" data-border-type="All" style="background-color: rgb(206, 182, 164); border-radius: 0px;"><div class="simblaEL containerHolder" data-drag="P29" id="P29" style="position: relative;"><div class="container" style="/* margin-top: 30px; *//* margin-bottom: 30px; */ border-radius: 0px;" data-border-type="All"><div class="row simblaEL rDivider" data-drag="P31" id="P31" style="position: relative; margin-top: 50px; margin-bottom: 70px;"><div class="sDivider col-md-12 index0" data-colsize="12"><div class="simblaEL tcH" data-drag="P33" id="P33" style="position: relative;"><div class="textContainer H1" style="cursor: text; margin-top: 0px; margin-bottom: 0px;"><h1 style="text-align: center;">OUR PROJECT</h1></div></div><div class="simblaEL tcH" data-drag="P35" id="P35"><div class="textContainer H3" style="cursor: text; line-height: 36px;"><h3 style="text-align: center; line-height: 36px;"><font color="#444444" style="font-size: 24px;">The goal of this project is to engineer a biological circuit utilizing multiple e.coli DNA regulatory sequences and a green fluorescent protein (GFP) reporter as a means to detect Arsenic. Once this construct is designed and constructed the circuit will be tested to demonstrate its viability using a cell free lateral flow paper based system designed to detect Arsenic.&nbsp;</font><div><br></div></h3></div></div></div></div><div class="row simblaEL rDivider" data-drag="P37" id="P37"><div class="sDivider col-md-12 index0" data-colsize="12"><div class="simblaEL simblaImg" data-drag="P39" id="P39"><img draggable="false" class="" alt="e8b9_arsenic_based_sea_monkeys_lab.jpg" data-id="" title="e8b9_arsenic_based_sea_monkeys_lab.jpg" src="https://media-cdn.tripadvisor.com/media/photo-s/07/d6/96/b6/downtown-tacoma-waterfront.jpg" width="200" height="600" ></div></div></div><div class="row simblaEL rDivider" data-drag="P41" id="P41" style="position: relative; margin-top: 40px;"><div class="sDivider col-md-3 index0" data-colsize="3"><div class="simblaEL tcH" data-drag="P49" id="P49" style="position: relative;"></div><div class="simblaEL tc" data-drag="P51" id="P51" style="position: relative;"></div><div class="simblaEL tcH" data-drag="P53" id="P53" style="position: relative;"></div><div class="simblaEL tcH" data-drag="P55" id="P55" style="position: relative;"></div><div class="simblaEL tc" data-drag="P57" id="P57" style="position: relative;"></div></div><div class="sDivider col-md-6 index1" data-colsize="6"><div class="simblaEL tcH" data-drag="P59" id="P59" style="position: relative;"><div class="textContainer H2" style="margin-left: 0px; cursor: text; padding-left: 20px; padding-top: 0px; line-height: 36px; margin-top: 0px;"><h2 style="text-align: left; line-height: 36px;">Resources</h2></div></div><div class="simblaEL separatorElm" data-drag="P61" id="P61" style="position: relative; padding-left: 10px; padding-right: 10px; margin-top: 30px; margin-bottom: 30px;"><div data-type="11" data-color="d3d3d3" style="width: 100%; padding-top: 0.6%; background-image: url(//sites.simbla.com/static-svg/11/d3d3d3); background-size: auto 100%;"></div></div><div class="simblaEL tc" data-drag="P63" id="P63" style="position: relative;"><div class="textContainer" style="cursor: text; padding: 10px 10px 30px; margin-top: 0px; margin-left: 0px; line-height: 36px;"><p style="text-align: left; line-height: 36px;"><font style="font-size: 24px;">Through this proof of concept we will be working closely with local Nonprofits Urban. The goal of this project is to engineer a biological circuit utilizing multiple e.coli DNA regulatory sequences and a green fluorescent protein (GFP) reporter as a means to detect Arsenic+.&nbsp;</font></p></div></div><div class="simblaEL tc" data-drag="P65" id="P65" style="position: relative;"><div class="textContainer" style="cursor: text; padding: 10px 10px 30px; margin-top: 0px; margin-left: 0px; line-height: 36px;"><p style="text-align: left; line-height: 36px;"><font style="font-size: 24px;">Once this construct is designed and constructed the goal will be to demonstrate this circuit as viable in a cell arsenic. Through this proof of concept we will be working closely with local Nonprofits Urban Waters and Readiness Acceleration and Innovation Network (RAIN) as well as a local high school Charles Wright Academy. Through this heave community involvement we hope to both increase pre-collegiate interest in STEM related fields, increase the communal understanding of the local environment and provide a cheap and effective tool for the detection of potentially toxic metals (Arsenic) and Readiness Acceleration and Innovation Network (RAIN) as well as a local high school Charles Wright Academy.</font></p></div></div><div class="simblaEL tcH" data-drag="P67" id="P67" style="position: relative;"></div><div class="simblaEL tc" data-drag="P69" id="P69" style="position: relative;"><div class="textContainer" style="cursor: text; padding: 10px 10px 30px; margin-top: 0px; margin-left: 0px; line-height: 36px;"><p style="text-align: left; line-height: 36px;"><font style="font-size: 24px;">&nbsp;Through this heave community involvement we hope to both increase pre-collegiate interest in STEM related fields, increase the communal understanding of the local environment and provide a cheap and effective tool for the detection of potentially toxic metals (Arsenic).
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<p><span id="docs-internal-guid-1b7da29c-fd54-c3a1-2826-3c55b3c31a0c"><span style="font-size: 12pt; color: rgb(0, 0, 0); background-color: transparent; vertical-align: baseline; white-space: pre-wrap;">The goal of this project is to engineer a biological circuit that utilizes </span><span style="font-size: 12pt; color: rgb(0, 0, 0); background-color: transparent; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 12pt; color: rgb(0, 0, 0); background-color: transparent; vertical-align: baseline; white-space: pre-wrap;"> plasmid DNA sequences and a green fluorescent protein (GFP) reporter to detect arsenic. The constitutive promoter in our construct produces a regulatory protein ArsR that binds with arsenic ions and triggers visible fluorescence. Our team will demonstrate this circuit using a cell-free lateral flow paper-based system. Ultimately, the purpose of this circuit will be to provide an inexpensive and ubiquitous arsenic sensor for testing environmental samples. </span></span>​</p>
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<h2 class="wsite-content-title" style="text-align:center;"><font size="5" color="#2a2a2a"><font>Project Description</font></font></h2>
 
  
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<div class="paragraph" style="text-align:center;"><font size="2">The goal of this project is to engineer a biological circuit utilizing multiple e.coli DNA regulatory sequences and a green fluorescent protein (GFP) reporter as a means to detect Arsenic (See following list of parts). Once this construct is designed and constructed the goal will be to demonstrate this circuit as viable in a cell free lateral flow paper based system for the detection of &nbsp;Arsenic. Through this proof of concept we will be working closely with local Nonprofits Urban WThe goal of this project is to engineer a biological circuit utilizing multiple e.coli DNA regulatory sequences and a green fluorescent protein (GFP) reporter as a means to detect Arsenic (See list of parts in parts section). Once this construct is designed and constructed the goal will be to demonstrate this circuit as viable in a cell arsenic. Through this proof of concept we will be working closely with local Nonprofits Urban Waters and Readiness Acceleration and Innovation Network (RAIN) as well as a local high school Charles Wright Academy. Through this heave community involvement we hope to both increase pre-collegiate interest in STEM related fields, increase the communal understanding of the local environment and provide a cheap and effective tool for the detection of potentially toxic metals (Arsenic) and Readiness Acceleration and Innovation Network (RAIN) as well as a local high school Charles Wright Academy. Through this heave community involvement we hope to both increase pre-collegiate interest in STEM related fields, increase the communal understanding of the local environment and provide a cheap and effective tool for the detection of potentially toxic metals (Arsenic).<br /></font><br /><span></span><br /></div>
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<div class="paragraph" style="text-align:center;">What is Synthetic Biology?<ul><li>Synthetic Biology is the shift from Understanding and modifying biology to treating biology as an engineering discipline where basic building blocks are assembled from the ground up into larger engineered systems<br /></li></ul><br /></div>
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<p><span style="font-size:20px;"><span id="docs-internal-guid-1b7da29c-09db-8e3c-11bc-a8b64c92574a"><span style="font-family: Arial; background-color: transparent; vertical-align: baseline; white-space: pre-wrap;">Design and create a synthetic biological circuit containing a reporter protein regulated by arsenic ions.</span></span></span></p>
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<div class="paragraph">Goals of Synthetic biology<ul><li>Develop tools to work at increasing levels of abstraction, and to enable facile manufacturing of designs</li><li>Leverage expertise from established engineering disciplines</li><li>Ultimately, forward engineer novel solutions to diverse problems</li></ul></div>
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<p><span style="line-height:1em;"><span style="font-size:16px;"><span style="letter-spacing: 1.1px; white-space: pre-wrap;">Arsenic was selected by Cadets2Vets as the first chemical we would attempt to detect using a biological circuit. It is naturally found in the environment, especially in groundwater, and around industrial plants as a byproduct of manufacturing practices.</span>​</span></span></p><p><br></p><p><span style="line-height:1em;"><span style="font-size:16px;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;">The detection of arsenic is especially important to team </span></span><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;">members in the Puget Sound area, because North Tacoma is a Superfund Site. The ASARCO smelting company smokestack, erected in 1917, was a symbol of economic prosperity and booming business in the developing city of Tacoma. The copper smelting site on Ruston’s shore in Puget Sound brought jobs and wealth to the area. Unfortunately, over the course of its 100 years of operation, the plant had been pumping toxic materials like arsenic and lead directly into the air, where it then settled into the surrounding soil and water of the greater Tacoma area.</span></span></span></span></p><p><br></p><p><span style="line-height:1em;"><span style="font-size:16px;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;">The smokestack was demolished in 1985, but there are still toxic levels of heavy metals in the soil over 30 years later. The city of Tacoma takes measures to warn new Tacoma homeowners about how to mitigate any increased levels of exposure to toxic materials in their soil, but must also take more drastic steps to protect users of publicly-owned land.</span><br class="kix-line-break"><br class="kix-line-break"><span style="vertical-align: baseline; white-space: pre-wrap;">The Department of Defense has a strong interest in developing rapid assays to test for environmental risks to protect the health of service members. Troops often deploy to hostile environments and assessing the risks of the environment itself can be challenging. The development of the arsenic circuit represents a new way to use synthetic biology in the military setting. Demonstrating that a biological circuit can be used in this application will lead to an expansion of the technology to test for more substances in the future. </span></span></span></span></p>
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<p><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;">The maximum contaminant level for arsenic that federal and state agencies use as guideline to test the arsenic levels in drinking water was established by the EPA. Initially it was set to 50 micrograms/liter or 50 ppb. However, drinking water that contains 50 ppb arsenic for prolonged periods of time may cause bladder or lung cancer and other chronic conditions.</span></span></span></span><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;"></span></span></span></span><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;"> </span></span></span></span><br></p><p><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;"></span></span></span></span><br></p><p><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;">In 2001, the EPA reduced the maximum contaminant level of arsenic to 10 micrograms/ liter or 10 ppb</span><span style="vertical-align: super; white-space: pre-wrap;">1</span><span style="vertical-align: baseline; white-space: pre-wrap;">. Nevertheless, according to data gathered between the years of 1993 and 2007, arsenic contamination of the waters in Washington state persisted</span><span style="vertical-align: super; white-space: pre-wrap;">2</span><span style="vertical-align: baseline; white-space: pre-wrap;">.</span></span></span></span><br></p><p><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;"></span></span></span></span><br></p><p><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;">Probes in the waters of western part of Washington state, </span></span></span></span><span style="font-size: 16px; letter-spacing: 1.1px; white-space: pre-wrap; background-color: rgb(255, 255, 255);">near the southern part of the state by the Columbia River, and also north near Everett</span><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;"> have detected higher concentrations of arsenic, close to 50 ppb. </span></span></span></span><br></p><p><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;"></span></span></span></span><br></p><p><span style="font-size:16px;"><span style="line-height:1em;"><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;">Developing an inexpensive paper-based circuit to detect the concentration of arsenic in the water in the region will greatly influence our knowledge and awareness of the quality of the water are consuming.</span></span><span style="letter-spacing: 1.1px;"><span style="vertical-align: baseline; white-space: pre-wrap;">We hope to develop our assay to detect arsenic within the EPA limit.</span></span></span></span></p>
 
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<p></p><ol><li><span><span style="font-size: 12pt; font-family: Arial; background-color: transparent; vertical-align: baseline; white-space: pre-wrap;">“Drinking Water Arsenic Rule History”, EPA.gov. https://www.epa.gov/dwreginfo/drinking-water-arsenic-rule-history. Web. Sept 06, 2017.</span></span><br></li><li><span><span id="docs-internal-guid-1b7da29c-fdbd-fb12-e772-bbf347097214"><span style="font-size: 12pt; font-family: Arial; background-color: transparent; vertical-align: baseline; white-space: pre-wrap;">“Arsenic Detection in Washington State Public Water Supplies”, doh.wa.gov. http://www.doh.wa.gov/CommunityandEnvironment/DrinkingWater/Contaminants/ArsenicinDrinkingWater/MapofArsenicDetections. Web. Sept 06, 2017.</span></span></span></li></ol>
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Latest revision as of 16:05, 1 November 2017

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Description - Home


DESCRIPTION OF OUR PROJECT



The goal of this project is to engineer a biological circuit that utilizes E. coli plasmid DNA sequences and a green fluorescent protein (GFP) reporter to detect arsenic. The constitutive promoter in our construct produces a regulatory protein ArsR that binds with arsenic ions and triggers visible fluorescence. Our team will demonstrate this circuit using a cell-free lateral flow paper-based system. Ultimately, the purpose of this circuit will be to provide an inexpensive and ubiquitous arsenic sensor for testing environmental samples. ​



Goal #1



Design and create a synthetic biological circuit containing a reporter protein regulated by arsenic ions.



Goal #2



Demonstrate that the circuit can detect arsenic in solution in cells or using cell-free lysates.



Goal #3



Determine the conditions needed for cell-free lysates to detect EPA limits for harmful concentrations of arsenic.



why we chose arsenic





Arsenic was selected by Cadets2Vets as the first chemical we would attempt to detect using a biological circuit. It is naturally found in the environment, especially in groundwater, and around industrial plants as a byproduct of manufacturing practices.​


The detection of arsenic is especially important to team members in the Puget Sound area, because North Tacoma is a Superfund Site. The ASARCO smelting company smokestack, erected in 1917, was a symbol of economic prosperity and booming business in the developing city of Tacoma. The copper smelting site on Ruston’s shore in Puget Sound brought jobs and wealth to the area. Unfortunately, over the course of its 100 years of operation, the plant had been pumping toxic materials like arsenic and lead directly into the air, where it then settled into the surrounding soil and water of the greater Tacoma area.


The smokestack was demolished in 1985, but there are still toxic levels of heavy metals in the soil over 30 years later. The city of Tacoma takes measures to warn new Tacoma homeowners about how to mitigate any increased levels of exposure to toxic materials in their soil, but must also take more drastic steps to protect users of publicly-owned land.

The Department of Defense has a strong interest in developing rapid assays to test for environmental risks to protect the health of service members. Troops often deploy to hostile environments and assessing the risks of the environment itself can be challenging. The development of the arsenic circuit represents a new way to use synthetic biology in the military setting. Demonstrating that a biological circuit can be used in this application will lead to an expansion of the technology to test for more substances in the future.



arsenic levels of interest





The maximum contaminant level for arsenic that federal and state agencies use as guideline to test the arsenic levels in drinking water was established by the EPA. Initially it was set to 50 micrograms/liter or 50 ppb. However, drinking water that contains 50 ppb arsenic for prolonged periods of time may cause bladder or lung cancer and other chronic conditions.


In 2001, the EPA reduced the maximum contaminant level of arsenic to 10 micrograms/ liter or 10 ppb1. Nevertheless, according to data gathered between the years of 1993 and 2007, arsenic contamination of the waters in Washington state persisted2.


Probes in the waters of western part of Washington state, near the southern part of the state by the Columbia River, and also north near Everett have detected higher concentrations of arsenic, close to 50 ppb.


Developing an inexpensive paper-based circuit to detect the concentration of arsenic in the water in the region will greatly influence our knowledge and awareness of the quality of the water are consuming.We hope to develop our assay to detect arsenic within the EPA limit.



Sources



  1. “Drinking Water Arsenic Rule History”, EPA.gov. https://www.epa.gov/dwreginfo/drinking-water-arsenic-rule-history. Web. Sept 06, 2017.
  2. “Arsenic Detection in Washington State Public Water Supplies”, doh.wa.gov. http://www.doh.wa.gov/CommunityandEnvironment/DrinkingWater/Contaminants/ArsenicinDrinkingWater/MapofArsenicDetections. Web. Sept 06, 2017.

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