http://2014.igem.org/wiki/index.php?title=Team:Washington/Future_Plans&feed=atom&action=historyTeam:Washington/Future Plans - Revision history2024-03-28T17:01:05ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014.igem.org/wiki/index.php?title=Team:Washington/Future_Plans&diff=390972&oldid=prevJhl325 at 02:43, 18 October 20142014-10-18T02:43:52Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <h1> Future Plans </h1></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <h1<ins class="diffchange diffchange-inline">><center</ins>> Future Plans <ins class="diffchange diffchange-inline"></center></ins></h1></div></td></tr>
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</table>Jhl325http://2014.igem.org/wiki/index.php?title=Team:Washington/Future_Plans&diff=383817&oldid=prevKristanguyen at 01:41, 18 October 20142014-10-18T01:41:50Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Like any new up and coming technique, the Degron system will require further testing with a larger variety of well studied mutant variants of a single protein, as well as a larger number of well studied proteins in general, before the system can truly be accepted. This is especially important because the proteins we have used to test the Degron system are mostly alpha-helical. In order to make sure that our system is generalizable to all protein topologies, we must test to see if we can obtain similar results with proteins of other topologies.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> Like any new up and coming technique, the Degron system will require further testing with a larger variety of well studied mutant variants of a single protein, as well as a larger number of well studied proteins in general, before the system can truly be accepted. This is especially important because the proteins we have used to test the Degron system are mostly alpha-helical. In order to make sure that our system is generalizable to all protein topologies, we must test to see if we can obtain similar results with proteins of other topologies. <ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
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</table>Kristanguyenhttp://2014.igem.org/wiki/index.php?title=Team:Washington/Future_Plans&diff=266736&oldid=prevJhl325: Future plans!2014-10-16T08:25:46Z<p>Future plans!</p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><h1> Future Plans </h1></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins><h1> Future Plans </h1></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h3> Examination of more proteins </h3></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h3> Examination of more proteins </h3></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins><p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>Like any new up and coming technique, the <del class="diffchange diffchange-inline">degron </del>system<del class="diffchange diffchange-inline">, </del>will require further testing with a larger variety of well studied mutant variants of a single protein as well as a larger number of well studied proteins in general before the system can truly be accepted. <del class="diffchange diffchange-inline"><br></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>Like any new up and coming technique, the <ins class="diffchange diffchange-inline">Degron </ins>system will require further testing with a larger variety of well studied mutant variants of a single protein<ins class="diffchange diffchange-inline">, </ins>as well as a larger number of well studied proteins in general<ins class="diffchange diffchange-inline">, </ins>before the system can truly be accepted. <ins class="diffchange diffchange-inline">This is especially important because </ins>the <ins class="diffchange diffchange-inline">proteins we have used to </ins>test <ins class="diffchange diffchange-inline">the Degron system </ins>are <ins class="diffchange diffchange-inline">mostly alpha-helical. In order </ins>to <ins class="diffchange diffchange-inline">make sure </ins>that <ins class="diffchange diffchange-inline">our </ins>system <ins class="diffchange diffchange-inline">is generalizable to all protein topologies</ins>, <ins class="diffchange diffchange-inline">we must test to see if we can obtain similar results with proteins </ins>of other <ins class="diffchange diffchange-inline">topologies</ins>.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> Our current plans for </del>the <del class="diffchange diffchange-inline">future are too </del>test <del class="diffchange diffchange-inline">a protein 33RM2 and its less stable variant 33CL1 both of which </del>are <del class="diffchange diffchange-inline">bind </del>to <del class="diffchange diffchange-inline">PD-1 (a negative t-cell regulator </del>that <del class="diffchange diffchange-inline">prevents the recognition of tumorous cells by the immune </del>system<del class="diffchange diffchange-inline">).</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> Since</del>, <del class="diffchange diffchange-inline">the stability </del>of <del class="diffchange diffchange-inline">both these proteins are known and have been verified using </del>other <del class="diffchange diffchange-inline">techniques such as thermal melts, they are very suitable candidates for testing using our degron system</del>. <br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> Our current plans for the future are to test a protein 33RM2 and its less stable variant, 33CL1, both of which bind to PD-1 (a negative t-cell regulator that prevents the recognition of tumorous cells by the immune system). Since the stability of both of these proteins are known and have been verified using other techniques, such as thermal melts, they are very suitable candidates for testing our Degron system.</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h3> Further evolving more stable variants of existing proteins </h3></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h3> Further evolving more stable variants of existing proteins </h3></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins><p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>Throughout this past summer, <del class="diffchange diffchange-inline">out </del>team has been evolving more stable variants of BINDI through error-prone PCR <del class="diffchange diffchange-inline">and going </del>forwards we will continue this process and continually analyze the mutants with flow cytometry and select cells that exhibit higher fluorescence with FACS.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>Throughout this past summer, <ins class="diffchange diffchange-inline">our </ins>team has been evolving more stable variants of BINDI through error-prone PCR<ins class="diffchange diffchange-inline">. Going </ins>forwards<ins class="diffchange diffchange-inline">, </ins>we will continue this process and <ins class="diffchange diffchange-inline">will </ins>continually analyze the mutants with flow cytometry and select cells that exhibit higher fluorescence with FACS.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> Furthermore, </del>our <del class="diffchange diffchange-inline">technique could be applied</del>...<br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> ***********NEEDS TO BE FINISHED*************** </del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </p></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <h3> Recommendations for Degron system use </h3></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <p></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> We offer a couple recommendations for those who would like to utilize </ins>our <ins class="diffchange diffchange-inline">novel method of protein stabilization:</ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> 1</ins>. <ins class="diffchange diffchange-inline">Use only Deg0, Deg1, and Deg2 for your stability testing</ins>. <ins class="diffchange diffchange-inline">We offer this recommendation because Deg3 and Deg4 are just a parallel version of Deg2 and Deg1, respectively</ins>. <ins class="diffchange diffchange-inline">That is, making all five constructs would be redundant as, theoretically, Deg1 and Deg4 (or Deg2 and Deg3) should have a similar stability, and hence GFP production. In order to minimize extra work, we recommend you utilize only Deg0, Deg1, and Deg2.</ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline">2. Utilize <i>E. coli</i> as your test organism, rather than <i>S. cervisiae </i>. Generally, <i> E. coli </i> is easier to work with as it has a faster growth and is very commonly used in laboratories.</ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <h3> Future Applications </h3></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> Protein engineering is highly important in many areas. Our system allows a generalizable and high throughput method of engineering proteins, and is also less time consuming than current stabilization methods.</ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> Engineered proteins selected through this method could be produced in bacteria and aid in the development of thermostable, de novo protein therapeutics.</ins></div></td></tr>
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</table>Jhl325http://2014.igem.org/wiki/index.php?title=Team:Washington/Future_Plans&diff=262775&oldid=prevKristanguyen: Created page with "{{Template:Team:UW/CSS}} <html> <body> <h1> Future Plans </h1> <h3> Examination of more proteins </h3> <p> Like any new up and coming techni..."2014-10-16T03:36:18Z<p>Created page with "{{Template:Team:UW/CSS}} <html> <body> <h1> Future Plans </h1> <h3> Examination of more proteins </h3> <p> Like any new up and coming techni..."</p>
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Like any new up and coming technique, the degron system, will require further testing with a larger variety of well studied mutant variants of a single protein as well as a larger number of well studied proteins in general before the system can truly be accepted. <br><br />
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Our current plans for the future are too test a protein 33RM2 and its less stable variant 33CL1 both of which are bind to PD-1 (a negative t-cell regulator that prevents the recognition of tumorous cells by the immune system).<br />
Since, the stability of both these proteins are known and have been verified using other techniques such as thermal melts, they are very suitable candidates for testing using our degron system. <br><br />
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<h3> Further evolving more stable variants of existing proteins </h3><br />
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Throughout this past summer, out team has been evolving more stable variants of BINDI through error-prone PCR and going forwards we will continue this process and continually analyze the mutants with flow cytometry and select cells that exhibit higher fluorescence with FACS.<br />
Furthermore, our technique could be applied...<br><br />
***********NEEDS TO BE FINISHED*************** <br />
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</html></div>Kristanguyen