Difference between revisions of "Team:Slovenia"

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         .igemSign {
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     $(document).ready(
 
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             function () {
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+
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                     $('img').each(function () {
 
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 +
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 +
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 +
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 +
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 +
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                 </a>
 
                 </a>
 
                 <div class="ui vertical sticky text menu">
 
                 <div class="ui vertical sticky text menu">
 +
                    <a class="item" href="//2016.igem.org/Team:Slovenia" style="color:#DB2828;">
 +
                        <i class="selected radio icon"></i>
 +
                        <b>Home</b>
 +
                    </a>
 
                     <a class="item" href="#intro" style="margin-left: 10%">
 
                     <a class="item" href="#intro" style="margin-left: 10%">
 
                         <i class="selected radio icon"></i>
 
                         <i class="selected radio icon"></i>
 
                         <b>Project</b>
 
                         <b>Project</b>
                     </a>
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                     </a>
 
                     <a class="item" href="#experts" style="margin-left: 10%">
 
                     <a class="item" href="#experts" style="margin-left: 10%">
 
                         <i class="selected radio icon"></i>
 
                         <i class="selected radio icon"></i>
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                     <span>
 
                     <span>
 
<br/>
 
<br/>
</span>
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</span>
                    <img onmouseenter="tooglePopup();" onmouseleave="tooglePopup();" class="ui large circular image"
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<div style = "position:fixed; width:14%; left:3%; bottom:3%;">
                        src="//2016.igem.org/wiki/images/c/cb/T--Slovenia--igemLogo.gif">
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<a href = "//2016.igem.org/Team:Slovenia/Proof"><img onmouseenter="tooglePopup();" onmouseleave="tooglePopup();" class="ui large circular image" src="//2016.igem.org/wiki/images/6/67/T--Slovenia--iGEM-gif.gif"></a>
                    <!-- <p style="font-size:11px;">
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<div class="popup igemSign">
                            The iGEM symbol was drawn with a glass rod letter by letter on engineered human cells and imaged by a camera.
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<span class="popuptext" id="igemSign">Touchpaint - detecting touch by light. The iGEM symbol was drawn with a glass rod letter by letter on engineered human cells and imaged by a camera.
                            Cells were transfected with constructs coding for the bacterial ion channel MscS, gas vesicles (GvpA and GvpC) and a Ca-dependent
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Cells were transfected with constructs coding for the bacterial ion channel MscS, gas vesicles (GvpA and GvpC) to enhance mechanosensing and a Ca-dependent
                            cyclic split luciferase reporter.
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                            cyclic split luciferase reporter to visualize the signal by light.</span>
                    </p> -->
+
</div>
                    <div class="popup igemSign">
+
</div>
<span class="popuptext" id="igemSign">The iGEM symbol was drawn with a glass rod letter by letter on engineered human cells and imaged by a camera.
+
                 </div>  
Cells were transfected with constructs coding for the bacterial ion channel MscS, gas vesicles (GvpA and GvpC) and a Ca-dependent
+
cyclic split luciferase reporter.</span>
+
                    </div>
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                 </div>
+
  
             </div>
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             </div>  
 
             <div class="article" id="context">
 
             <div class="article" id="context">
 
                 <!-- menu goes here -->
 
                 <!-- menu goes here -->
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                 <div class="main ui citing justified container">
 
                 <div class="main ui citing justified container">
 
                     <div>
 
                     <div>
                         <h1 class="ui centered dividing header"><span id="intro" class="section"> &nbsp; </span>Sonicell
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                         <h1 class="ui centered dividing header" style="font-size: 3rem"><span id="intro" class="section colorize"> &nbsp; </span>Sonicell
 
                         </h1>
 
                         </h1>
 
                         <div class="ui segment">
 
                         <div class="ui segment">
                             <p>Project Sonicell introduces exciting foundational advances to synthetic biology aimed to
+
                             <p>For the therapy of diseases such as diabetes, we require a system of drug delivery that is fast, controllable and noninvasive. Engineering cells to produce drugs in the tissue is a promising direction. However, in most genetic circuits designed by synthetic biology the response is delayed due to the need to transcripe and translate the mediators. Project Sonicell introduces exciting foundational advances to synthetic biology aimed to
 
                                 enable rapid cellular response to a combination of external stimuli such as sound, light
 
                                 enable rapid cellular response to a combination of external stimuli such as sound, light
                                 or chemical compounds. This system is composed of a module for enhanced sensitivity of
+
                                 or chemical compounds. This system is composed of a module for <b>enhanced sensitivity of
                                 cells to ultrasound or other mechanical stimuli sensed by a calcium-dependent reporter,
+
                                 cells to ultrasound or other mechanical stimuli</b>, sensed by a calcium-dependent reporter,
                                 and a module for integration of a combination of several input signals into a signaling
+
                                 and a module for integration of a combination of several input signals into a <b>signaling
                                 pathway based on the collection of orthogonal proteases. Finally, the proteases were
+
                                 pathway based on proteolysis </b> by a collection of orthogonal proteases. Finally, the proteases were
 
                                 designed to cleave an endoplasmic reticulum retention signal from target proteins, which
 
                                 designed to cleave an endoplasmic reticulum retention signal from target proteins, which
                                 results in a secretion of premade proteins.</p>
+
                                 results in <b>secretion of premade proteins</b>.</p>
 
                             <div>
 
                             <div>
 
                                 <div class="container">
 
                                 <div class="container">
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                     </div>
 
                     </div>
 
                     <div class="ui segment">
 
                     <div class="ui segment">
                         <h4><span id="experts" class="section">&nbsp;</span>Abstract for experts</h4>
+
                         <h4><span id="experts" class="section colorize">&nbsp;</span>Abstract for experts</h4>
 
                         <p>Synthetic biology opens exciting perspectives to control cells, for applications ranging from
 
                         <p>Synthetic biology opens exciting perspectives to control cells, for applications ranging from
 
                             industrial processes to cell-based therapy. However, the large majority of designed cellular
 
                             industrial processes to cell-based therapy. However, the large majority of designed cellular
                             circuits are based on the transcriptional regulation, which may be too slow for many
+
                             circuits are based on transcriptional regulation, which may be too slow for many
 
                             therapeutic or diagnostic applications, for example delivery of insulin or detection of a
 
                             therapeutic or diagnostic applications, for example delivery of insulin or detection of a
 
                             metabolite. Several medical doctors and researchers that we consulted stressed that a fast
 
                             metabolite. Several medical doctors and researchers that we consulted stressed that a fast
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                             several advantages of light with the added ability to penetrate tissue.</p>
 
                             several advantages of light with the added ability to penetrate tissue.</p>
 
                         <p>In our project we enhanced the sensitivity of mammalian cells to ultrasound or other
 
                         <p>In our project we enhanced the sensitivity of mammalian cells to ultrasound or other
                             mechanical stimulus by introduction of bacterial or engineered mammalian mechanosensors.
+
                             mechanical stimuli by introduction of bacterial or engineered mammalian mechanosensors.
 
                             Additionally, the response to ultrasound and touch was strongly increased by expression of
 
                             Additionally, the response to ultrasound and touch was strongly increased by expression of
 
                             the two components of bacterial gas vesicles, GvpA and GvpC. Mechanosensing was detected by
 
                             the two components of bacterial gas vesicles, GvpA and GvpC. Mechanosensing was detected by
 
                             the calcium-induced calmodulin-M13 complex reconstituting split cyclic luciferase, highly
 
                             the calcium-induced calmodulin-M13 complex reconstituting split cyclic luciferase, highly
 
                             applicable for the emerging field of mechanogenetics. This enabled us to draw on cells using
 
                             applicable for the emerging field of mechanogenetics. This enabled us to draw on cells using
                             touch, where we engaged in collaboration with an artist.</p>
+
                             touch, where we engaged in collaboration with the artist Laura Olalde.</p>
 
                         <p>For the rapid response of cells to multiple stimuli we designed proteolysis-based signaling
 
                         <p>For the rapid response of cells to multiple stimuli we designed proteolysis-based signaling
                             pathways. For this purpose four orthogonal split proteases were generated, each recognizing
+
                             pathways. Four orthogonal split proteases were generated, each recognizing
 
                             its own motif of seven amino acid residues. Based on cleavage of coiled-coil dimerizing
 
                             its own motif of seven amino acid residues. Based on cleavage of coiled-coil dimerizing
 
                             domains we demonstrated the ability to implement proteolysis-based signal pathways and logic
 
                             domains we demonstrated the ability to implement proteolysis-based signal pathways and logic
 
                             functions in mammalian cells. Based on the cleavage of an ER retention peptide by a
 
                             functions in mammalian cells. Based on the cleavage of an ER retention peptide by a
                             protease, input signals lead to protein secretion without the slow step of induced protein
+
                             protease, input signals led to protein secretion without the slow step of induced protein
 
                             synthesis.</p>
 
                             synthesis.</p>
 
                         <p>We believe that this project introduced several foundational advances that could be very
 
                         <p>We believe that this project introduced several foundational advances that could be very
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                     </div>
 
                     </div>
 
                     <div class="ui segment">
 
                     <div class="ui segment">
                         <h4><span id="plain" class="section">&nbsp;</span>Abstract in plain English</h4>
+
                         <h4><span id="plain" class="section colorize">&nbsp;</span>Abstract in plain English</h4>
 
                         <p>
 
                         <p>
 
                             Synthetic biology aims to control cells so they can obey our commands and do what we want,
 
                             Synthetic biology aims to control cells so they can obey our commands and do what we want,
                             for example to produce drugs when needed. In our project we made cells respond to ultrasound
+
                             for example produce drugs when needed. In our project we made cells which respond to ultrasound
 
                             or touch. When we touch the cells they light up, which can be recorded on a camera. Ideally
 
                             or touch. When we touch the cells they light up, which can be recorded on a camera. Ideally
 
                             we want cells to respond to our commands as fast as possible, because sometimes we can’t
 
                             we want cells to respond to our commands as fast as possible, because sometimes we can’t
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                     </div>
 
                     </div>
 
                     <div>
 
                     <div>
                         <h1 class="ui centered dividing header"><span id="achievements" class="section"> &nbsp; </span>Achievements
+
                         <h1 class="ui centered dividing header"><span id="achievements" class="section colorize"> &nbsp; </span>Achievements
 
                         </h1>
 
                         </h1>
 
                         <div class="ui segment">
 
                         <div class="ui segment">
 
                             <div class="corners" style="float:right;">
 
                             <div class="corners" style="float:right;">
 
                                 <p><img src="//2016.igem.org/wiki/images/d/dc/T--Slovenia--starSmall.png"
 
                                 <p><img src="//2016.igem.org/wiki/images/d/dc/T--Slovenia--starSmall.png"
                                         alt="newAtiGEM" width="28" height="28" style="display: inline;"> new at science
+
                                         alt="newAtiGEM" width="28" height="28" style="display: inline;"> new in science
 
                                 </p>
 
                                 </p>
 
                                 <p><img src="//2016.igem.org/wiki/images/0/0d/T--Slovenia--igemLogoSmall.png"
 
                                 <p><img src="//2016.igem.org/wiki/images/0/0d/T--Slovenia--igemLogoSmall.png"
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                                             src="//2016.igem.org/wiki/images/d/dc/T--Slovenia--starSmall.png"
 
                                             src="//2016.igem.org/wiki/images/d/dc/T--Slovenia--starSmall.png"
 
                                             alt="newAtiGEM" width="28" height="28" style="display: inline;"></li>
 
                                             alt="newAtiGEM" width="28" height="28" style="display: inline;"></li>
                                 <li>A circular proteolysis-activated luciferase reporter was experimentally verified and
+
                                 <li>A cyclic proteolysis-activated luciferase reporter was experimentally verified and
 
                                     introduced into the iGEM collection <img
 
                                     introduced into the iGEM collection <img
 
                                             src="//2016.igem.org/wiki/images/0/0d/T--Slovenia--igemLogoSmall.png"
 
                                             src="//2016.igem.org/wiki/images/0/0d/T--Slovenia--igemLogoSmall.png"
 
                                             alt="newAtiGEM" width="30" height="30" style="display: inline;"></li>
 
                                             alt="newAtiGEM" width="30" height="30" style="display: inline;"></li>
 
                                 <li>A set of four different orthogonal site-specific proteases was designed and tested
 
                                 <li>A set of four different orthogonal site-specific proteases was designed and tested
                                     as split proteins in mammalian cells <img
+
                                     as split proteins with induced reconstitution in mammalian cells <img
 
                                             src="//2016.igem.org/wiki/images/d/dc/T--Slovenia--starSmall.png"
 
                                             src="//2016.igem.org/wiki/images/d/dc/T--Slovenia--starSmall.png"
 
                                             alt="newAtiGEM" width="28" height="28" style="display: inline;"></li>
 
                                             alt="newAtiGEM" width="28" height="28" style="display: inline;"></li>
                                 <li>New orthogonal protease-based signaling pathways and information processing platform
+
                                 <li>New orthogonal protease-based signaling pathways were designed as an information     processing platform
                                    was designed and several logic functions based on the combination of multiple input
+
                                  and several logic functions based on the combination of multiple input
 
                                     signals were tested experimentally <img
 
                                     signals were tested experimentally <img
 
                                             src="//2016.igem.org/wiki/images/d/dc/T--Slovenia--starSmall.png"
 
                                             src="//2016.igem.org/wiki/images/d/dc/T--Slovenia--starSmall.png"
 
                                             alt="newAtiGEM" width="28" height="28" style="display: inline;"></li>
 
                                             alt="newAtiGEM" width="28" height="28" style="display: inline;"></li>
                                 <li>Proteolysis of the ER retention signal was introduced as the trigger for the fast
+
                                 <li>Proteolysis of ER retention signal was introduced as the trigger for the fast
 
                                     release of proteins from cells aimed to enable fast therapeutic responses such as
 
                                     release of proteins from cells aimed to enable fast therapeutic responses such as
 
                                     required for the release of peptide hormones, neuroactive peptides etc. <img
 
                                     required for the release of peptide hormones, neuroactive peptides etc. <img
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                     </div>
 
                     </div>
 
                     <div>
 
                     <div>
                         <h1 class="ui centered dividing header"><span id="requirements" class="section"> &nbsp; </span>Medal
+
                         <h1 class="ui centered dividing header"><span id="requirements" class="section colorize"> &nbsp; </span>Medal
 
                             requirements</h1>
 
                             requirements</h1>
 +
                        <div class="ui basic segment">
 
                             <table class="ui collapsing unstackable celled table" style="box-shadow: 0 1px 2px 0 rgba(34,36,38,.15)">
 
                             <table class="ui collapsing unstackable celled table" style="box-shadow: 0 1px 2px 0 rgba(34,36,38,.15)">
 
                                 <thead class="full-width">
 
                                 <thead class="full-width">
 
                                 <tr class="center aligned">
 
                                 <tr class="center aligned">
 
                                     <th> Medal</th>
 
                                     <th> Medal</th>
                                    <th> Explanation</th>
 
 
                                     <th> Criteria</th>
 
                                     <th> Criteria</th>
 +
                                    <th> Explanation</th>
 
                                     <th></th>
 
                                     <th></th>
 
                                 </tr>
 
                                 </tr>
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                                 <tbody>
 
                                 <tbody>
 
                                 <tr>
 
                                 <tr>
                                     <td colspan="4" class="center aligned" style="background-color: #CD7F32">BRONZE</td>
+
                                     <td colspan="4" class="center aligned" style="background-color: gold">GOLD</td>
 
                                 </tr>
 
                                 </tr>
 
                                 <tr>
 
                                 <tr>
                                     <td>Register and attend</td>
+
                                     <td>Integrated Human Practices</td>
                                     <td>Register for iGEM.</td>
+
                                     <td>Expand on your silver medal activity by demonstrating how you have integrated
                                     <td>We have successfully registered.</td>
+
                                        the investigated issues into
 +
                                        the design and/or execution of your project.
 +
                                    </td>
 +
                                     <td>Implementation of several <a
 +
                                            href="https://2016.igem.org/Team:Slovenia/Integrated_Practices">experts from
 +
                                        different medical fields and culturologists</a> helped us
 +
                                        improve our
 +
                                        project.
 +
                                    </td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                 </tr>
 
                                 </tr>
 
                                 <tr>
 
                                 <tr>
                                     <td>Deliverables</td>
+
                                     <td>Improve a previous part or project
                                     <td>Meet all deliverables on the Requirements page.</td>
+
                                    </td>
                                     <td>We met all the listed deliverables.</td>
+
                                     <td>Improve the function OR characterization of an existing BioBrick Part or Device
 +
                                        and enter this information
 +
                                        in the Registry.
 +
                                    </td>
 +
                                     <td>We <a href="https://2016.igem.org/Team:Slovenia/Parts">improved</a> the parts
 +
                                        <a href="http://parts.igem.org/Part:BBa_K737005:Experience">BBa_K737005</a> and <a
 +
                                                href="http://parts.igem.org/Part:BBa_K157010:Experience">BBa_K157010</a>
 +
                                        by equipping them with additional tags, expressing them
 +
                                        in human cells and further characterizing their function.
 +
                                    </td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                 </tr>
 
                                 </tr>
 
                                 <tr>
 
                                 <tr>
                                     <td>Attribution</td>
+
                                     <td>Proof of concept</td>
                                     <td>Create a page on your team wiki with clear attribution of each aspect of your
+
                                     <td>Demonstrate a functional proof of concept of your project. </td>
                                        project.
+
                                     <td>We successfully demonstrated the functionality of selected (split) proteases and
                                    </td>
+
                                        used them to <a
                                     <td>We created a <a href="//2016.igem.org/Team:Slovenia">wiki page</a> describing
+
                                                href="https://2016.igem.org/Team:Slovenia/Demonstrate">control
                                        the attributions to the
+
                                            secretion</a> of the reporter protein from the cell.
                                        project.
+
 
                                     </td>
 
                                     </td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                 </tr>
 
                                 </tr>
 
                                 <tr>
 
                                 <tr>
                                     <td>Part / Contribution</td>
+
                                     <td>Demonstrate your work</td>
                                     <td>Document at least one new standard BioBrick Part or Device central to your
+
                                     <td>How your project works under real-world conditions.
                                        project and submit this part to
+
                                        the iGEM Registry.
+
 
                                     </td>
 
                                     </td>
                                     <td>We documented and submitted 45 standard <a
+
                                     <td>We showed that our mechanosensing constructs coexpressed in human cells allow
                                            href="//parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2016&group=Slovenia">BioBrick
+
                                        for a controlled response to
                                        parts</a>.
+
                                        touch and used them for our art application, <a
 +
                                                href="//2016.igem.org/Team:Slovenia/Proof">Touch painting</a>.
 
                                     </td>
 
                                     </td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                     <td><i class="large green checkmark icon"></i></td>
Line 436: Line 468:
 
                                     <td>We demonstrated the functionality of our constructs and provided experimental
 
                                     <td>We demonstrated the functionality of our constructs and provided experimental
 
                                         data. We created a list of our
 
                                         data. We created a list of our
                                         <a href="https://2016.igem.org/Team:Slovenia/Favorite_Part">favorite parts</a>
+
                                         <a href="https://2016.igem.org/Team:Slovenia/Parts">favorite parts</a>
 
                                         and detailed our experiences with them.
 
                                         and detailed our experiences with them.
 
                                     </td>
 
                                     </td>
Line 473: Line 505:
 
                                 </tr>
 
                                 </tr>
 
                                 <tr>
 
                                 <tr>
                                     <td colspan="4" class="center aligned" style="background-color: gold">GOLD</td>
+
                                     <td colspan="4" class="center aligned" style="background-color: #CD7F32">BRONZE</td>
 
                                 </tr>
 
                                 </tr>
 
                                 <tr>
 
                                 <tr>
                                     <td>Integrated Human Practices</td>
+
                                     <td>Register and attend</td>
                                     <td>Expand on your silver medal activity by demonstrating how you have integrated
+
                                     <td>Register for iGEM.</td>
                                        the investigated issues into
+
                                     <td>We have successfully registered.</td>
                                        the design and/or execution of your project.
+
                                    </td>
+
                                     <td>Implementation of several <a
+
                                            href="https://2016.igem.org/Team:Slovenia/Integrated_Practices">experts from
+
                                        different medical fields and culturologists</a> helped us
+
                                        improve our
+
                                        project.
+
                                    </td>
+
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                 </tr>
 
                                 </tr>
 
                                 <tr>
 
                                 <tr>
                                     <td>Improve a previous part or project
+
                                     <td>Deliverables</td>
                                    </td>
+
                                     <td>Meet all deliverables on the Requirements page.</td>
                                     <td>Improve the function OR characterization of an existing BioBrick Part or Device
+
                                     <td>We met all the listed deliverables.</td>
                                        and enter this information
+
                                        in the Registry.
+
                                    </td>
+
                                     <td>We <a href="https://2016.igem.org/Team:Slovenia/Parts">improved</a> the parts
+
                                        <a href="http://parts.igem.org/Part:BBa_K1965003">BBa_K1965003</a> and <a
+
                                                href="http://parts.igem.org/Part:BBa_K1965030">BBa_K1965030</a>
+
                                        by equipping them with additional tags, expressing them
+
                                        in human cells and further characterizing their function.
+
                                    </td>
+
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                 </tr>
 
                                 </tr>
 
                                 <tr>
 
                                 <tr>
                                     <td>Proof of concept</td>
+
                                     <td>Attribution</td>
                                     <td>Demonstrate a functional proof of concept of your project. </td>
+
                                     <td>Create a page on your team wiki with clear attribution of each aspect of your
                                     <td>We successfully demonstrated the functionality of selected (split) proteases and
+
                                        project.
                                        used them to <a
+
                                    </td>
                                                href="https://2016.igem.org/Team:Slovenia/Demonstrate">control
+
                                     <td>We created a <a href="//2016.igem.org/Team:Slovenia/Attributions">wiki page</a> describing
                                            secretion</a> of the reporter protein from the cell.
+
                                        the attributions to the
 +
                                        project.
 
                                     </td>
 
                                     </td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                 </tr>
 
                                 </tr>
 
                                 <tr>
 
                                 <tr>
                                     <td>Demonstrate your work</td>
+
                                     <td>Part / Contribution</td>
                                     <td>How your project works under real-world conditions.
+
                                     <td>Document at least one new standard BioBrick Part or Device central to your
 +
                                        project and submit this part to
 +
                                        the iGEM Registry.
 
                                     </td>
 
                                     </td>
                                     <td>We showed that our mechanosensing constructs coexpressed in human cells allow
+
                                     <td>We documented and submitted 52 standard <a
                                        for a controlled response to
+
                                            href="//parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2016&group=Slovenia">BioBrick
                                        touch and used them for our art application, <a
+
                                        parts</a>.
                                                href="//2016.igem.org/Team:Slovenia/Proof">Touch painting</a>.
+
 
                                     </td>
 
                                     </td>
 
                                     <td><i class="large green checkmark icon"></i></td>
 
                                     <td><i class="large green checkmark icon"></i></td>
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                                 </tbody>
 
                             </table>
 
                             </table>
 +
                        </div>
 
                     </div>
 
                     </div>
 
                 </div>
 
                 </div>
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     </div>
 
     </div>
 
</div>
 
</div>
 +
<div>
 +
<a href="//igem.org/Main_Page">
 +
<img border="0" alt="iGEM" src="//2016.igem.org/wiki/images/8/84/T--Slovenia--logo_250x250.png" width="5%" style = "position: fixed; bottom:0%; right:1%;">
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Latest revision as of 13:50, 19 October 2016

Home

  Sonicell

For the therapy of diseases such as diabetes, we require a system of drug delivery that is fast, controllable and noninvasive. Engineering cells to produce drugs in the tissue is a promising direction. However, in most genetic circuits designed by synthetic biology the response is delayed due to the need to transcripe and translate the mediators. Project Sonicell introduces exciting foundational advances to synthetic biology aimed to enable rapid cellular response to a combination of external stimuli such as sound, light or chemical compounds. This system is composed of a module for enhanced sensitivity of cells to ultrasound or other mechanical stimuli, sensed by a calcium-dependent reporter, and a module for integration of a combination of several input signals into a signaling pathway based on proteolysis by a collection of orthogonal proteases. Finally, the proteases were designed to cleave an endoplasmic reticulum retention signal from target proteins, which results in secretion of premade proteins.

project scheme module1 module1 module1 module1

 Abstract for experts

Synthetic biology opens exciting perspectives to control cells, for applications ranging from industrial processes to cell-based therapy. However, the large majority of designed cellular circuits are based on transcriptional regulation, which may be too slow for many therapeutic or diagnostic applications, for example delivery of insulin or detection of a metabolite. Several medical doctors and researchers that we consulted stressed that a fast but controllable response is high on their wish list of expectations from synthetic biology. Additionally, noninvasive stimulation of selected tissues in the organism would also be highly desirable. While light is extremely useful as a rapid, spatially-restricted input signal, it cannot penetrate deep into the tissue. On the other hand, ultrasound combines several advantages of light with the added ability to penetrate tissue.

In our project we enhanced the sensitivity of mammalian cells to ultrasound or other mechanical stimuli by introduction of bacterial or engineered mammalian mechanosensors. Additionally, the response to ultrasound and touch was strongly increased by expression of the two components of bacterial gas vesicles, GvpA and GvpC. Mechanosensing was detected by the calcium-induced calmodulin-M13 complex reconstituting split cyclic luciferase, highly applicable for the emerging field of mechanogenetics. This enabled us to draw on cells using touch, where we engaged in collaboration with the artist Laura Olalde.

For the rapid response of cells to multiple stimuli we designed proteolysis-based signaling pathways. Four orthogonal split proteases were generated, each recognizing its own motif of seven amino acid residues. Based on cleavage of coiled-coil dimerizing domains we demonstrated the ability to implement proteolysis-based signal pathways and logic functions in mammalian cells. Based on the cleavage of an ER retention peptide by a protease, input signals led to protein secretion without the slow step of induced protein synthesis.

We believe that this project introduced several foundational advances that could be very useful to synthetic biology far beyond iGEM and for the benefit of humanity for therapy, diagnostics and potentially many other advanced applications.

 Abstract in plain English

Synthetic biology aims to control cells so they can obey our commands and do what we want, for example produce drugs when needed. In our project we made cells which respond to ultrasound or touch. When we touch the cells they light up, which can be recorded on a camera. Ideally we want cells to respond to our commands as fast as possible, because sometimes we can’t wait an hour before the cells produce the medicine and release it. That is why we gave cells a novel mechanism of processing information. We achieved this by combining several enzymes that recognize very specific parts of proteins and cut them, which changes their function. This allowed us to combine different signals, like sound, touch, light or chemicals, to obtain the desired cell response. The new enzymes can also cut the anchor with which medicines are attached to cells after the cells make them. Among many possible uses of our inventions, we can imagine activating cells in the brain by ultrasound, which means that we don’t need to use surgery to help people with Parkinson’s disease, or can trigger fast production of insulin in the body, to help people with diabetes.

  Achievements

newAtiGEM new in science

newAtiGEM new at iGEM

  • Mammalian cell sensitivity to ultrasound and mechanical stimuli was increased by ectopic expression of bacterial or human cation permeable channels and functional reconstitution of bacterial protein gas vesicles from two protein components (GvpA and GvpC) newAtiGEM
  • A custom-made ultrasound generator device was used to stimulate mammalian cells newAtiGEM
  • A mechano-sensory luciferase reporter sensitive to an influx of free calcium ions was introduced into mammalian cells, which enabled rapid light emission of mammalian cells in response to mechanical stimuli and enabled painting on cells by touch with exciting potentials for other applications newAtiGEM
  • A cyclic proteolysis-activated luciferase reporter was experimentally verified and introduced into the iGEM collection newAtiGEM
  • A set of four different orthogonal site-specific proteases was designed and tested as split proteins with induced reconstitution in mammalian cells newAtiGEM
  • New orthogonal protease-based signaling pathways were designed as an information processing platform and several logic functions based on the combination of multiple input signals were tested experimentally newAtiGEM
  • Proteolysis of ER retention signal was introduced as the trigger for the fast release of proteins from cells aimed to enable fast therapeutic responses such as required for the release of peptide hormones, neuroactive peptides etc. newAtiGEM

  Medal requirements

Medal Criteria Explanation
GOLD
Integrated Human Practices Expand on your silver medal activity by demonstrating how you have integrated the investigated issues into the design and/or execution of your project. Implementation of several experts from different medical fields and culturologists helped us improve our project.
Improve a previous part or project Improve the function OR characterization of an existing BioBrick Part or Device and enter this information in the Registry. We improved the parts BBa_K737005 and BBa_K157010 by equipping them with additional tags, expressing them in human cells and further characterizing their function.
Proof of concept Demonstrate a functional proof of concept of your project. We successfully demonstrated the functionality of selected (split) proteases and used them to control secretion of the reporter protein from the cell.
Demonstrate your work How your project works under real-world conditions. We showed that our mechanosensing constructs coexpressed in human cells allow for a controlled response to touch and used them for our art application, Touch painting.
SILVER
Validated Part / Validated Contribution Experimentally validate that at least one new BioBrick Part or Device of your own design and construction works as expected. We demonstrated the functionality of our constructs and provided experimental data. We created a list of our favorite parts and detailed our experiences with them.
Collaboration Convince the judges you have helped any registered iGEM team from high school, a different track, another university, or another institution in a significant way. During our project we had several skype meetings with other iGEM teams. We also provided iGEM Team biotINK from Munich with BBa_K782063 created by team Slovenia 2012.
Human Practices Demonstrate how your team has identified, investigated, and addressed one or more of issues (education, public engagement, public policy issues, public perception, or other activities) in the context of your project. Education and transmission of interest in science is an important part of our project. This is why we prepared several lectures for high school students and also collaborated with an artist who gave our project a new perspective by conveying science to public through art.
BRONZE
Register and attend Register for iGEM. We have successfully registered.
Deliverables Meet all deliverables on the Requirements page. We met all the listed deliverables.
Attribution Create a page on your team wiki with clear attribution of each aspect of your project. We created a wiki page describing the attributions to the project.
Part / Contribution Document at least one new standard BioBrick Part or Device central to your project and submit this part to the iGEM Registry. We documented and submitted 52 standard BioBrick parts.