Team:William and Mary/project

From 2014.igem.org

(Difference between revisions)

Latest revision as of 20:25, 17 October 2014

Project

Click on a blueprint to progress to the next phase of construction!

OUR GOAL: The goal of this project was to create a set of tools that would open up the door to studying calcium dynamics to future teams. Calcium is a crucial signal in many aspects for eukaryotic organisms, including brain development and function; however, creating molecular tools to study calcium dynamics in eukaryotes is extremely difficult. By creating parts to emulate, measure, and record calcium spikes in model prokaryotes, our team hoped to pave the way for future teams to build genetic circuits for eukaryotic systems. We planned to create three different classes of parts that can: 1. Create calcium influx in bacteria
2. Indicate the presence of calcium
3. Keep track of calcium spikes and produce a reporter molecule after a certain number of spikes

Click on the construction site to progress to the next phase!

3. Tracking and reporting Ca2+ spikes:

A calcium-dependent promoter would activate a genetic counting circuit that would, after a specified number of spikes, produce a reporter gene to indicate that this number of spikes occurred in the cells.

1. Creating Ca2+ influx in bacteria:
The green algae Chlamydomonas naturally produces two non-specific cation channels that activate in response to particular wavelengths of light. Named channelrhodopsin-1 (ChR1) and channelrhodopsin-2 (ChR2), we thought that these could be used to induce calcium spikes in prokaryotes. There exist multiple varieties of ChR1 and ChR2, which have been optimized for different ions and conditions. 2. Indicating presence of Ca2+:
Genetically Encoded Calcium Indicators (GECIs) are fusion proteins of a fluorescent protein (XFP) and calmodulin (calcium-binding messenger). XFP is initially in a non-fluorescing state. Upon binding of Ca2+, a conformational change in calmodulin activates the XFP, producing fluorescent light.

Future Directions: With the tools that have been developed and are continuing to be developed, the William & Mary iGEM team is making huge amounts of progress towards achieving our goals of inducing calcium spikes, reporting the presence of calcium, and counting calcium spikes in prokaryotes. We have adapted two channelrhodopsins to make them usable by iGEM teams, and have made great strides towards having a functioning RCaMP (or GCaMP) available to be used in future iGEM projects. We hope that opening up this door will create more options for future teams intending to study this important signaling molecule.

The Calcium Kids (and our marvelous PI!)

Click on the photo to look at our Parts page!

@@ Line 1: / Line 1: @@
-<!DOCTYPE html>
 <html class="html" lang="en-US">
   <head>
+<style media="screen" type="text/css">
+#content{
+    width: 100%;
+}
+#globalWrapper{
+    padding-bottom: 0px;
+}
+/* Removes header footer and borders */
+#contentSub, #search-controls, .firstHeading, #footer-box, #catlinks, #p-logo {
+    display:none;}
+#top-section {
+    border: none;
+    height: 0px;}
+#content {
+    border: none;}
+/* Removes "teams" from the menubar */
+#menubar.left-menu.noprint > ul > li:last-child {
+    display: none;}
+/* Resizes the menubar to fik the links (default is 400px) */
+#menubar {
+    width: auto;}
+body {
+    margin: 10px 0 0 0;
+    padding: 0;}
+#top-section {
+    width: 965px;
+    height: 0;
+    margin: 0 auto;
+    padding: 0;
+    border: none;}
+#menubar {
+    font-size: 65%;
+    top: -14px;}
+.left-menu:hover {
+    background-color: transparent;}
+#menubar li a {
+    background-color: transparent;}
+#menubar:hover {
+    color: white;}
+#menubar li a {
+    color: transparent;}
+#menubar:hover li a {
+    color: white;}
+</style>
    <script type="text/javascript">
@@ Line 11: / Line 62: @@
    <title>Project</title>
    <!-- CSS -->
-   <link rel="stylesheet" type="text/css" href="css/site_global.css?518065832"/>
+   <link rel="stylesheet" type="text/css" href="https://2014.igem.org/Team:William_and_Mary/wm_global.css?action=raw&amp;ctype=text/css"/>
-   <link rel="stylesheet" type="text/css" href="css/master_a-master.css?25009745"/>
+   <link rel="stylesheet" type="text/css" href="https://2014.igem.org/Team:William_and_Mary/wm_master.css?action=raw&amp;ctype=text/css"/>
-   <link rel="stylesheet" type="text/css" href="css/project.css?3804403314" id="pagesheet"/>
+   <link rel="stylesheet" type="text/css" href="https://2014.igem.org/Team:William_and_Mary/wm_project.css?action=raw&amp;ctype=text/css" id="pagesheet"/>
    <!-- Other scripts -->
    <script type="text/javascript">
@@ Line 32: / Line 83: @@
         <div class="clearfix" id="u841_align_to_page">
          <a class="nonblock nontext clip_frame grpelem" id="u1083" href="https://2014.igem.org/Main_Page"><!-- image --><img class="block" id="u1083_img" src="https://static.igem.org/mediawiki/2014/c/c2/WM_Igem_logo_circle.png" alt="" width="150" height="150"/></a>
-         <a class="nonblock nontext grpelem" id="u842-4" href="https://2014.igem.org/Team:William_and_Mary"><!-- rasterized frame --><img id="u842-4_img" alt="The Calcium Kit" width="537" height="73" src="images/u842-4.png"/></a>
+         <a class="nonblock nontext grpelem" id="u842-4" href="https://2014.igem.org/Team:William_and_Mary"><!-- rasterized frame --><img id="u842-4_img" alt="The Calcium Kit" width="537" height="73" src="https://static.igem.org/mediawiki/2014/4/45/WM_Header_CalciumKit_U842-4.png"/></a>
          <a class="nonblock nontext clip_frame grpelem" id="u874" href="https://igem.org/Team.cgi?year=2014&amp;team_name=William_and_Mary"><!-- image --><img class="block" id="u874_img" src="https://static.igem.org/mediawiki/2014/6/6b/WM_Constructioncolif.png" alt="" width="168" height="134"/></a>
         </div>
@@ Line 38: / Line 89: @@
       </div>
       <div class="clip_frame grpelem" id="u846"><!-- image -->
-       <img class="block" id="u846_img" src="images/williamandmarywolf.png" alt="" width="532" height="84"/>
+       <img class="block" id="u846_img" src="https://static.igem.org/mediawiki/2014/e/ec/Williamandmarywolf.png" alt="" width="532" height="84"/>
       </div>
       <a class="anchor_item grpelem" id="top"></a>
@@ Line 44: / Line 95: @@
      <nav class="MenuBar clearfix colelem" id="menuu386"><!-- horizontal box -->
       <div class="MenuItemContainer clearfix grpelem" id="u408"><!-- vertical box -->
-       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u411" href="https://2014.igem.org/Team:William_and_Mary"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u413" alt="Home" src="images/blank.gif"/><!-- state-based BG images --></a>
+       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u411" href="https://2014.igem.org/Team:William_and_Mary"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u413" alt="Home" src="https://static.igem.org/mediawiki/2014/a/a0/WM_Nav_Home_U413.png"/><!-- state-based BG images --></a>
       </div>
       <div class="MenuItemContainer clearfix grpelem" id="u526"><!-- vertical box -->
-       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u529" href="https://2014.igem.org/Team:William_and_Mary/team"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u531" alt="Team" src="images/blank.gif"/><!-- state-based BG images --></a>
+       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u529" href="https://2014.igem.org/Team:William_and_Mary/team"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u531" alt="Team" src="https://static.igem.org/mediawiki/2014/8/82/WM_Nav_Team_U531.png"/><!-- state-based BG images --></a>
       </div>
       <div class="MenuItemContainer clearfix grpelem" id="u446"><!-- vertical box -->
-       <a class="nonblock nontext MenuItem MenuItemWithSubMenu MuseMenuActive clearfix colelem" id="u449" href="https://2014.igem.org/Team:William_and_Mary/project"><!-- horizontal box --><div class="MenuItemLabel NoWrap grpelem" id="u451-4"><!-- rasterized frame --><div id="u451-4_clip"><img id="u451-4_img" alt="Project" width="135" height="29" src="images/u451-4-a.png"/></div></div></a>
+       <a class="nonblock nontext MenuItem MenuItemWithSubMenu MuseMenuActive clearfix colelem" id="u449" href="https://2014.igem.org/Team:William_and_Mary/project"><!-- horizontal box --><div class="MenuItemLabel NoWrap grpelem" id="u451-4"><!-- rasterized frame --><div id="u451-4_clip"><img id="u451-4_img" alt="Project" width="135" height="29" src="https://static.igem.org/mediawiki/2014/5/57/WM_Nav_Project_U451-4-a.png"/></div></div></a>
       </div>
       <div class="MenuItemContainer clearfix grpelem" id="u394"><!-- vertical box -->
-       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u397" href="https://2014.igem.org/Team:William_and_Mary/parts"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u400" alt="Parts" src="images/blank.gif"/><!-- state-based BG images --></a>
+       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u397" href="https://2014.igem.org/Team:William_and_Mary/parts"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u400" alt="Parts" src="https://static.igem.org/mediawiki/2014/5/50/WM_Nav_Parts_U400.png"/><!-- state-based BG images --></a>
       </div>
       <div class="MenuItemContainer clearfix grpelem" id="u401"><!-- vertical box -->
-       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u404" href="https://2014.igem.org/Team:William_and_Mary/notebook"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u406" alt="Notebook" src="images/blank.gif"/><!-- state-based BG images --></a>
+       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u404" href="https://2014.igem.org/Team:William_and_Mary/notebook"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u406" alt="Notebook" src="https://static.igem.org/mediawiki/2014/1/1c/WM_Nav_Notebook_U406.png"/><!-- state-based BG images --></a>
       </div>
       <div class="MenuItemContainer clearfix grpelem" id="u387"><!-- vertical box -->
-       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u388" href="https://2014.igem.org/Team:William_and_Mary/safety"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u389" alt="Safety" src="images/blank.gif"/><!-- state-based BG images --></a>
+       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u388" href="https://2014.igem.org/Team:William_and_Mary/safety"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u389" alt="Safety" src="https://static.igem.org/mediawiki/2014/b/b5/WM_Nav_Safety_U389.png"/><!-- state-based BG images --></a>
       </div>
       <div class="MenuItemContainer clearfix grpelem" id="u925"><!-- vertical box -->
-       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u926" href="https://2014.igem.org/Team:William_and_Mary/thanks"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u929" alt="Thanks" src="images/blank.gif"/><!-- state-based BG images --></a>
+       <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u926" href="https://2014.igem.org/Team:William_and_Mary/thanks"><!-- horizontal box --><img class="MenuItemLabel NoWrap grpelem" id="u929" alt="Thanks" src="https://static.igem.org/mediawiki/2014/0/04/WM_Nav_Thanks_U929.png"/><!-- state-based BG images --></a>
       </div>
      </nav>
@@ Line 74: / Line 125: @@
       <img class="grpelem" id="u645-20" alt="OUR GOAL: The goal of this project was to create a set of tools that would open up the door to studying calcium dynamics to future teams. Calcium is a crucial signal in many aspects for eukaryotic organisms, including brain development and function; however, creating molecular tools to study calcium dynamics in eukaryotes is extremely difficult. By creating parts to emulate, measure, and record calcium spikes in model prokaryotes, our team hoped to pave the way for future teams to build genetic circuits for eukaryotic systems. We planned to create three different classes of parts that can: 1. Create calcium influx in bacteria
 . Indicate the presence of calcium
-. Keep track of calcium spikes and produce a reporter molecule after a certain number of spikes " width="587" height="612" src="images/u645-20.png"/><!-- rasterized frame -->
+. Keep track of calcium spikes and produce a reporter molecule after a certain number of spikes " width="587" height="612" src="https://static.igem.org/mediawiki/2014/5/53/WM_Project_Goal-blurb_U645-20.png"/><!-- rasterized frame -->
      </div>
      <div class="clearfix colelem" id="pu646-3"><!-- group -->
@@ Line 101: / Line 152: @@
        </div>
       </div>
-      <img class="grpelem" id="u894-4" alt="Groundwork:" width="313" height="42" src="images/u894-4.png"/><!-- rasterized frame -->
+      <img class="grpelem" id="u894-4" alt="Groundwork:" width="313" height="42" src="https://static.igem.org/mediawiki/2014/a/a3/WM_Project_Groundwork_U894-4.png"/><!-- rasterized frame -->
       <img class="grpelem" id="u896-15" alt="&nbsp;1. Creating Ca2+ influx in bacteria:
 The green algae Chlamydomonas naturally produces two non&#45;specific cation channels that activate in response to particular wavelengths of light. Named channelrhodopsin&#45;1 (ChR1) and channelrhodopsin&#45;2 (ChR2), we thought that these could be used to induce calcium spikes in prokaryotes. There exist multiple varieties of ChR1 and ChR2, which have been optimized for different ions and conditions. 2. Indicating presence of Ca2+:
-Genetically Encoded Calcium Indicators (GECIs) are fusion proteins of a fluorescent protein (XFP) and calmodulin (calcium&#45;binding messenger). XFP is initially in a non&#45;fluorescing state. Upon binding of Ca2+, a conformational change in calmodulin activates the XFP, producing fluorescent light. " width="384" height="759" src="images/u896-15.png"/><!-- rasterized frame -->
+Genetically Encoded Calcium Indicators (GECIs) are fusion proteins of a fluorescent protein (XFP) and calmodulin (calcium&#45;binding messenger). XFP is initially in a non&#45;fluorescing state. Upon binding of Ca2+, a conformational change in calmodulin activates the XFP, producing fluorescent light. " width="384" height="759" src="https://static.igem.org/mediawiki/2014/a/a4/WM_Project_Groundwork_U896-15.png"/><!-- rasterized frame -->
      </div>
      <div class="clearfix colelem" id="pfuture_plans"><!-- group -->
       <a class="anchor_item grpelem" id="future_plans"></a>
-      <img class="grpelem" id="u683-7" alt="Future Directions: With the tools that have been developed and are continuing to be developed, the William &amp; Mary iGEM team is making huge amounts of progress towards achieving our goals of inducing calcium spikes, reporting the presence of calcium, and counting calcium spikes in prokaryotes. We have adapted two channelrhodopsins to make them usable by iGEM teams, and have made great strides towards having a functioning RCaMP (or GCaMP) available to be used in future iGEM projects. We hope that opening up this door will create more options for future teams intending to study this important signaling molecule." width="812" height="314" src="images/u683-7.png"/><!-- rasterized frame -->
+      <img class="grpelem" id="u683-7" alt="Future Directions: With the tools that have been developed and are continuing to be developed, the William &amp; Mary iGEM team is making huge amounts of progress towards achieving our goals of inducing calcium spikes, reporting the presence of calcium, and counting calcium spikes in prokaryotes. We have adapted two channelrhodopsins to make them usable by iGEM teams, and have made great strides towards having a functioning RCaMP (or GCaMP) available to be used in future iGEM projects. We hope that opening up this door will create more options for future teams intending to study this important signaling molecule." width="812" height="314" src="https://static.igem.org/mediawiki/2014/1/1c/WM_Project_FutureDirections-blurb_U683-7.png"/><!-- rasterized frame -->
      </div>
-     <a class="nonblock nontext clip_frame colelem" id="u677" href="https://2014.igem.org/Team:William_and_Mary/parts"><!-- image --><img class="block" id="u677_img" src="images/team_isc.jpg" alt="" width="582" height="873"/></a>
+     <a class="nonblock nontext clip_frame colelem" id="u677" href="https://2014.igem.org/Team:William_and_Mary/parts"><!-- image --><img class="block" id="u677_img" src="https://static.igem.org/mediawiki/2014/0/0f/Team_isc.jpg" alt="" width="582" height="873"/></a>
      <div class="verticalspacer"></div>
      <div class="clearfix colelem" id="u684-6"><!-- content -->