Team:SJTU-BioX-Shanghai

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                    <li><a href="/Team:SJTU-BioX-Shanghai/Project">Project</a>
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Overview
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|Various methods have been developed to regulate gene expressions on different levels so far. Though widely used in recent study, these methods are difficult to be applied on multiple genes simultaneously and independently due to the complex procedures or the high cost. Therefore, we aim to build a versatile system to control and regulate different genes in an easy and accurate way.
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|Thus we choose several light-controlled gene expression systems and a newly devised tool named CRISPRi to find the solution towards this problem. By changing light intensity, we hope to regulate different genes to different certain expressing levels, thus maximizing producing efficiency in a metabolic pathway. Light sensors are widely used in engineered biosynthetic parts for their sensibility and adjustable intensity, and CRISPRi can knock down different genes easily and specificly by changing the short sequence of its sgRNA part. With advantages of these two systems combined together, parameter determination of multiple gene-expressions will be more predictable and precise in our project, and this novel device will provide us with a promising way of genome-scale metabolism regulation.
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|align="center"|[[Team:SJTU-BioX-Shanghai | Team SJTU-BioX-Shanghai]]
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!align="center"|[[Team:SJTU-BioX-Shanghai|Home]]
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!align="center"|[[Team:SJTU-BioX-Shanghai/Team|Team]]
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!align="center"|[https://igem.org/Team.cgi?year=2013&team_name=SJTU-BioX-Shanghai Official Team Profile]
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!align="center"|[[Team:SJTU-BioX-Shanghai/Project|Project]]
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<div id="abstract-title"><p><strong style="color:rgb(255,102,102); font-size:60px;">A</strong>bstract</p></div>
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<p>Few researches have been done to regulate gene expression levels in genomic scale so far. This year we aim to combine two systems together in order to provide a universal and convenient tool which can be used to regulate different genomic genes simultaneously and independently in a quantitative way. </p>
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!align="center"|[[Team:SJTU-BioX-Shanghai/Safety|Safety]]
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<p>Our project involves the newly developed gene regulating tool CRISPRi and three light-controlled expression systems induced by red, green, and blue light respectively. Simply by changing the regulating parts in CRISPRi system towards mRFP, luciferase, and three enzymes, we hope to prove our system can be used qualitatively, quantitatively and practically step by step. </p>
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!align="center"|[[Team:SJTU-BioX-Shanghai/Attributions|Attributions]]
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<p>We have also designed a box and written a software as our experiment measurements. Simply by typing in several parameters, different gene expression levels can be controlled. This system can also be improved to predict the maximized producing efficiency after some simple tests in future.  </p>
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                                <p>Luminous Device provide us a tool to easily adjust genetic expression. With this tool, we can also regulate 3 genes at the same time.</p>
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                                <p>Use date collected from experiments, we can optimize the metabolic system after several circles.</p>
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<p><strong><img src="/wiki/images/7/73/12SJTU_r.png"><font face="Cambria, serif " size="4"> The honors we won:</font></strong> Gold Medal, Best New BioBrick Part or Device Engineered (BBa_K771001) and Regional Winner at Asia Jamboree; Advanced to World Championship.</p>
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<p><strong><font face="Wingdings " size="4">ü</font><font face="Cambria, serif " size="4"> The breakthrough we made:</font></strong> Redefinition of scaffold in Synthetic Biology by recruiting <i>E.coli’</i>s inner membrane as a natural two-dimensional scaffold. </p>
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<p><strong><font face="Wingdings " size="4">ü</font><font face="Cambria, serif " size="4"> The system we built:</font></strong> 6 membrane proteins orderly organized on the inner membrane of <i>E.coli</i>, the efficiency of which has been proved by fluorescence complementation assay and biosynthesis experiment.</p>
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<p><strong><font face="Wingdings " size="4">ü</font><font face="Cambria, serif " size="4"> The device we created – Membrane Accelerator:</font></strong> A universal tool that serves to accelerate biochemical reactions in <i>E.coli</i>; Rate of fatty acids synthesis was increased by 24 fold compared to wild-type <i>E.coli </i>and 9 fold compared to that with overexpressed cytoplasmic enzymes.</p>
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<p><strong><font face="Wingdings " size="4">ü</font><font face="Cambria, serif " size="4"> device we created – Membrane Rudder:</font></strong> A universal tool used to dynamically and artificially control biochemical reactions in <i>E.coli</i>; the direction of Violacein and Deoxyviolacein synthetic pathway was successfully switched. </p>
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<p><strong><font face="Wingdings " size="4">ü</font><font face="Cambria, serif " size="4"> New direction we proposed:</font></strong> The application of scaffold system in accelerating biodegradation pathway using our Membrane Accelerator.</p>
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<p><strong><font face="Wingdings " size="4">ü</font><font face="Cambria, serif " size="4"> Parts we submitted:</font></strong> 42 well-characterized parts that could either be used directly or serve as a universal tool readily for potential scientific or engineering use.</p>
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<p><strong><font face="Wingdings " size="4">ü</font><font face="Cambria, serif " size="4"> A club we established – BioCraft:</font></strong> The headquarter of our human practice programs, having come a long way in propagandizing Synthetic Biology and iGEM. Warmly-received activities have been held in and outside the campus. Several celebrities in different fields have shown support for us, laying a cornerstone for our future development.</p>
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Revision as of 13:43, 25 September 2013

Abstract

Few researches have been done to regulate gene expression levels in genomic scale so far. This year we aim to combine two systems together in order to provide a universal and convenient tool which can be used to regulate different genomic genes simultaneously and independently in a quantitative way.

Our project involves the newly developed gene regulating tool CRISPRi and three light-controlled expression systems induced by red, green, and blue light respectively. Simply by changing the regulating parts in CRISPRi system towards mRFP, luciferase, and three enzymes, we hope to prove our system can be used qualitatively, quantitatively and practically step by step.

We have also designed a box and written a software as our experiment measurements. Simply by typing in several parameters, different gene expression levels can be controlled. This system can also be improved to predict the maximized producing efficiency after some simple tests in future.

Luminous Device provide us a tool to easily adjust genetic expression. With this tool, we can also regulate 3 genes at the same time.

Use date collected from experiments, we can optimize the metabolic system after several circles.

The honors we won: Gold Medal, Best New BioBrick Part or Device Engineered (BBa_K771001) and Regional Winner at Asia Jamboree; Advanced to World Championship.

ü The breakthrough we made: Redefinition of scaffold in Synthetic Biology by recruiting E.coli’s inner membrane as a natural two-dimensional scaffold.

ü The system we built: 6 membrane proteins orderly organized on the inner membrane of E.coli, the efficiency of which has been proved by fluorescence complementation assay and biosynthesis experiment.

ü The device we created – Membrane Accelerator: A universal tool that serves to accelerate biochemical reactions in E.coli; Rate of fatty acids synthesis was increased by 24 fold compared to wild-type E.coli and 9 fold compared to that with overexpressed cytoplasmic enzymes.

ü device we created – Membrane Rudder: A universal tool used to dynamically and artificially control biochemical reactions in E.coli; the direction of Violacein and Deoxyviolacein synthetic pathway was successfully switched.

ü New direction we proposed: The application of scaffold system in accelerating biodegradation pathway using our Membrane Accelerator.

ü Parts we submitted: 42 well-characterized parts that could either be used directly or serve as a universal tool readily for potential scientific or engineering use.

ü A club we established – BioCraft: The headquarter of our human practice programs, having come a long way in propagandizing Synthetic Biology and iGEM. Warmly-received activities have been held in and outside the campus. Several celebrities in different fields have shown support for us, laying a cornerstone for our future development.


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