Team:TCU Taiwan/SystemDesign

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     <td width="128" height="128" align="left" valign="middle"><img src="https://static.igem.org/mediawiki/2014/4/43/TCU_Gear-128.png" width="128" height="128"></td>
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     <td width="128" height="128" align="left" valign="middle"><img src="https://static.igem.org/mediawiki/2014/4/4d/TCU_Blueprint-128.png" width="128" height="128"></td>
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     <td width="68%" align="left" valign="middle"><h6><font face="Trebuchet MS" size="7" color="#484644">CRISPR</font></h6></td>
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     <td width="68%" align="left" valign="middle"><h6><font face="Trebuchet MS" size="7" color="#484644">System Design</font></h6></td>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Team"><font size="3">Members</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Team"><font size="3">Members</font></a></li>
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      <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Achievements"><font size="3">Achievements</font></a></li>
       <li><a href="https://igem.org/Team.cgi?id=1473" target="_blank"><font size="3">Official Team Profile</font></a></li>
       <li><a href="https://igem.org/Team.cgi?id=1473" target="_blank"><font size="3">Official Team Profile</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Contact"><font size="3">Contact</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Contact"><font size="3">Contact</font></a></li>
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   <li><a href="#" style="cursor:default"><font size="5" style="color:#DBAC52">Project</font></a>
   <li><a href="#" style="cursor:default"><font size="5" style="color:#DBAC52">Project</font></a>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Project" style="color:#DBAC52"><font size="3">Introduction</font></a></li>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Project"><font size="3">Introduction</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Parts"><font size="3">Parts</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Parts"><font size="3">Parts</font></a></li>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/SystemDesign"><font size="3">System Design</font></a></li>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/SystemDesign"  style="color:#DBAC52"><font size="3">System Design</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/M13Phage"><font size="3">M13 Phage</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/M13Phage"><font size="3">M13 Phage</font></a></li>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/CRISPR"  style="color:#DBAC52"><font size="3">CRISPR</font></a></li>
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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/CRISPR"><font size="3">CRISPR</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/FutureWorks"><font size="3">Future Works</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/FutureWorks"><font size="3">Future Works</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/HumanPratics"><font size="3">Human Pratics</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/HumanPratics"><font size="3">Human Pratics</font></a></li>
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      <td colspan="3" style="background-color:#FFF2B5" height="20"><font face="Trebuchet MS" size="6" color="#A66B38">CRISPR Construction</font></td>
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                <td><img src="https://static.igem.org/mediawiki/2014/d/d8/TCU_SD10733136_390371157783708_1318933891_o.jpg" width="100%"/></td>
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                <td align="center"><font size="3" face="Verdana"><strong>Fig.1</strong></font></td>
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            <td width="15">&nbsp;</td>
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            <td><font size="3" face="Verdana" color="#333"><p>We synthesised a new CIRSPR system based on  a CRISPR designed by Standford-Brown 2013 (<a href="http://parts.igem.org/Part:BBa_K1218011" target="_blank">BBa_K1218011</a>). Here we  analyzed its sequence and then removed its Cas9 protein with Cas9&rsquo;s promoter  because we want our Cas9&rsquo;s expression can be inducible. So then we synthesized  a new  Cas9 structure which contains an  inducible promoter, pRha (<a href="http://parts.igem.org/Part:BBa_K914003" target="_blank">BBa_K914003</a>).  This promoter can be induced by rhamnose and silenced by glucose, both sugar  can be find in nature. Next, we put our inducible Cas9 back to the CRISPR  system, and because tracrRNA and crRNA have no function without Cas9 protein,  this CRISPR is now under our control.</p></font></td>
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        <td colspan="3" style="background-color:#FFF2B5"><font face="Trebuchet MS" size="6" color="#A66B38">Recombinant Phagemid Construction</font></td>
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            <td valign="top"><p><font color="#333" size="3" face="Verdana">After constructing our CRISPR system, we  ligated it with a red fluorescent reporter gene (<a href="http://parts.igem.org/Part:BBa_I13521" target="_blank">BBa_I13521</a>), so once our  system is spread, all bacterium get this system will become red. Finally, we  put whole system into phagemid pBluescript II SK(-), and get the recombinant  phagemid for M13 infection.</font></p></td>
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                <td><img src="https://static.igem.org/mediawiki/2014/4/4b/TCU_SD_10572757_390371167783707_1137689428_o.jpg" width="100%" /></td>
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                <td align="center"><font size="3" face="Verdana"><strong>Fig.2</strong></font></td>
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Latest revision as of 20:57, 17 October 2014


 
System Design
  
 
CRISPR Construction
     
Fig.1
 

We synthesised a new CIRSPR system based on a CRISPR designed by Standford-Brown 2013 (BBa_K1218011). Here we analyzed its sequence and then removed its Cas9 protein with Cas9’s promoter because we want our Cas9’s expression can be inducible. So then we synthesized a new  Cas9 structure which contains an inducible promoter, pRha (BBa_K914003). This promoter can be induced by rhamnose and silenced by glucose, both sugar can be find in nature. Next, we put our inducible Cas9 back to the CRISPR system, and because tracrRNA and crRNA have no function without Cas9 protein, this CRISPR is now under our control.
 
Recombinant Phagemid Construction
     

After constructing our CRISPR system, we ligated it with a red fluorescent reporter gene (BBa_I13521), so once our system is spread, all bacterium get this system will become red. Finally, we put whole system into phagemid pBluescript II SK(-), and get the recombinant phagemid for M13 infection.

  
Fig.2
 
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Team Members Project Parts Human Pratics Modeling Safety Notebook Attributions

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