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Newer edit →

Line 266:

content.childs=[35];

content.titleShort = "pDawn";

-

content.titleLong = "";

+

content.titleLong = "pDawn system"; (langer Titel)

-

content.summary= "";

+

content.summary= "The plasmid pDawn was designed by Ohlendorf et al.<sup><a href=#ref21.1>21.1</a></sup> in 2012 together with its counter plasmid pDusk. Both plasmids are single plasmid systems, which allow the activation (pDawn) or repression (pDusk) of gene expression by blue light. They are easy to implement in the laboratory and lead to up to 460-fold activity change upon ilumination.";

-

content.text= "";

+

content.text= "The plasmid pDawn was designed by Ohlendorf et al.<sup><a href=#ref21.1>21.1</a></sup> in 2012 together with its counter plasmid pDusk. Both plasmids are single plasmid systems, which allow the activation (pDawn) or repression (pDusk) of gene expression by blue light. They described their system as easy to implement in the laboratory and suitable for both preparative and analytical purposes. Further, they were able to show up to 460-fold induction upon ilumination using their system. </br></br> Because, pDawn (and pDusk) is a system that regulates gene expression and thus acts on DNA level it is an eligible candidate for us to compare the speed of action of our system, which acts on protein level to an DNA regulation system. As an additional benefit, with pDawn, we implement a powerful light-induceable system, with low background expression and high dark/light expression difference in the iGEM database. </br></br> Ohlendorf et al. (2012)<sup><a href=#ref21.1>21.1</a></sup> had previously designed the histidine kinase YF1, which phosphorylates FixJ in the absence of blue light. The phosphorylated FixJ drives robust gene expression from the FixK2 promotor as shown in figure 1 for both pDawn and the pDusk. Upon light absorption, net kinase activity of YF1 and consequently gene expression under control of the FixK2 promotor is greatly reduced. pDusk an pDawn enable light-repressed or light-induced gene expression in Escherichia coli and are easy to implement in the laboratory. Therefore, all components of the YF1/FixJ TCS were assembled on only one medium-copy plasmid in which YF1 and FixJ are constitutively expressed by the Lac<sup>q</sup> promotor. Target genes can be introduced via the multiple-cloning site (MCS) under the control of the pFixK2 promotor, allowing light-repressed gene expression. To obtain pDawn the light effect is inverted by the introduction of a gene-inversion cassette into pDusk. This gene-inversion cassette is based on the λ phage repressor cI and the λ phage promotor pR. (see figure 1)</br></br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/a/a1/Bonn_MS_Figure1_pDusk%26pDawn.jpg></br>Figure 1: Plasmids pDusk an pDawn. YF1 and FixJ are constitutively expressed by the Lacq promotor. Target genes can be introduced via the multiple-cloning site (MCS) under the control of the pFixK2 promotor. (Ohlendorf et al. 2012<sup><a href=#ref21.1>21.1</a></sup>)</div></br>Ohlendorf et al. (2012)<sup><a href=#ref21.1>21.1</a></sup> analyzed the system by the insertion of the red-fluorescent reporter protein DsRed Express2. That way they were able to measure up to 460-fold induction upon ilumination. However, they also found that their plasmid, which acts on gene expression level, only allows light-induced perturbations on the timescale of hours.</br></br><h2>References</h2></br><p><a name=ref21.1>21.1</a> <a href=http://www.sciencedirect.com/science/article/pii/S0022283612000113> Ohlendorf et al. (2009) From Dusk till Dawn One-Plasmid Systems for Light-Regulated Gene Expression.</a></p>";

content.type="Background";

break;

Line 316:

+

case 36:

content.i = 36;

-

content.parents=[19];

+

content.parents=[37, 19];

-

content.childs=[37];

+

content.childs=[];

-

content.titleShort = "LOV-ipaA - VinD1";

+

content.titleShort = "LOV-ipaA & VinD1";

-

content.titleLong = "";

+

content.titleLong = "LOV-ipaA & VinD1";

-

content.summary= "";

+

content.summary= "The LOV-ipaA -vinculin system is a combined system for light inducible heterodimerisation. This powerful tool, which allows photocontroled complex formation was establish by Lungu et al. in 2012.";

-

content.text= "";

+

content.text= "The LOV-ipaA -vinculin system is a combined system for light inducible heterodimerisation. It consists out of a LOV domain, which undergoes conformational changes upon irradiation with blue light, and the ipaA-vinculin hybridization system. This two building blocks have be combined and described by Lungu et al. in 2012.</br></br> Lungu et al. (2008) where able to measure a 49-fold difference in target binding upon irradiation as compared to the dark state. However, they further modified the system by mutations of the LOV-ipaA construct and successfully weakend the baseline affinity for vinculin (initial design: 3.5 nM to 69 nM; mutant: 2.4 nM to >40µM affinity for vinculin) to reduce the dark state activity. </br></br> Lov-ipaA-VinD1 is a powerful tool which allows photocontroled complex formation. To establish this system Lungu et al. (2012)<sup><a href=#ref36.1>36.1</a></sup> fused the Ja helix of the LOV Domain to ipaA.</br>To be more precise they used the LOV2 domain from Avena sativa photopropin 1 (AsLOV2), which – as previously shown – can be used to photomodulate the affinity of peptides for their binding partners (see Figure 1). </br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/0/02/Bonn_MS_Figure1_LOV.jpg></br>Figure 1: General design of AsLOV2 fusion proteins (Lungu et al. 2012)<sup><a href=#ref36.1>36.1</a></sup></div> </br>In other studies had been shown that the LOV domain can be fused to entire protein domains, allowing photomodulation of the protein binding. However, they stated that it might be of high importance to bring the LOV domain closer to ipaA, in order to allow photomodulation in this case, because ipaA is only a peptide and thus more flexible than folded domains.</br></br>Therefore, Lungu et al. (2012)<sup><a href=#ref36.1>36.1</a></sup> identified similar amino acid sequences in the ipaA peptide and the Ja helix of the LOV Domain and used this combined with molecular modeling to create photomodulateable AsLOV2-ipaA (see Figure 2). </br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/5/54/Bonn_MS_Figure2_LOV-ipaA.jpg></br>Figure 2: Light-inducible LOV-ipaA construct (Lungu et al. 2012)<sup><a href=#ref36.1>36.1</a></sup></div></br>They were able to proof the functionality of the AsLOV2-ipaA system by heterodimerization in yeast (yeast two-hybrid system or Y2H) The yeast two-hybrid system can be used to monitor protein–protein interactions between two proteins. The system is based on a transcription factor, which is split into two separate fragments, called the binding domain (BD) and activating domain (AD). Each domain is fused to one protein and thus only if the proteins interact with each other BD and AD are close enough to initiate the transcription of a reporter gene.</br></br>The basic principle of the LOV-ipaA & VinD1 system works as follows. In the dark state the fusion product LOV-ipaA is not able to form a complex with vinculin, because LOV blocks ipaA sterically. However, activation of the LOV domain with blue light induces conformational changes in the fused molecule, which results in a movement of the Ja helix with the ipaA away from LOV. Thereby, ipaA becomes accessible for VinD1 and a Complex is formed.</br></br>The activation is reversible and the entire system can be genetically encoded. This two facts are the main advantages of this system in contrast to other typically used systems, which like for the chemical system for example, are based on in vivo covalently modified peptides, that can be activated by light induced cleavage. Moreover, the protein used are relatively small and thus should interfere as little as possible with the prokaryotic metabolism, the activity change form dark to light state is high, the system is completely genetically encoded and reversible. But, the most important property of this system is that it allows the light-controlled heterodimerisation of the two split variants of sspB, which is necessary for our system.</br></br><h2>References</h2></br><a name=ref36.1>36.1</a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3334866/>Lungu et al. (2012) Designing photoswitchable peptides using the AsLOV2 domain</a>";

content.type="Background";

break;

Line 638:

Line 639:

break;

+

case 55:

+

content.i = 55;

+

content.parents=[53];

+

content.childs=[];

+

content.titleShort = "Protein function";

+

content.titleLong = "Analysis of protein function";

+

content.summary= "To understand the role of a specific gene or DNA region is one of the big challenges in lifescience research. Our system, which allows the fast and convenient elimination of defined proteins, is a new improved technique, with many advantages.";

+

content.text= "To understand the role of a specific gene or DNA region is one of the big challenges in lifescience research. Our system, which allows the fast and convenient elimination of defined proteins, is a new improved technique, with many advantages. It allows the control of protein activity with respect to time and space, is fast, robust, can be used for different proteins and changes the protein sequence only little, because the ssrA tag consist only out of 15 amino acids. Recently technologies like gene knockout or knockdown had been developed, which allow to investigate the role of a particular gene or DNA region by comparing the knockout organism to a wildtype with a similar genetic background. </br></br> A knockout means that a particular gene is deleted from the genome of an organism. This organism might be bacteria or yeast, but also eukaryotic cells, plants or even animals. To create a knockout organism recombinant DNA is inserted into a gene (Bartke, 2006<sup><a href=#ref55.3>55.3</a></sup>). When a genes sequence is interrupted, it may still be translated, but the resulting protein will be nonfunctional. Moreover, it is possible to knockout the gene only in defined tissues or at defined time points. This technique is called a conditional knockout. </br></br> On the other hand the knockdown, does not eliminate the specific gene on DNA, but on RNA level. Here interfering RNAs (siRNA) are inserted into the cell, leading to the degradation of the genes mRNA (Pratt and MacRae, 2009<sup><a href=#ref55.1>55.1</a></sup>) and hence no protein can be produced. </br></br> In contrast to the knockout and knockdown, our system allows the expression of the gene and the translation into functional protein. However, irradiation with blue light leads to the fast elimination of the particular protein. Therefore, one of the major advantages of our system is its speed, not only in comparison to knockout and knockdown, but also in comparison to other protein level systems. Comparing our system, to other protein level systems, like the system developed by Davis et al.<sup><a href=#ref55.2>55.2</a></sup> in 2011, which are induced by small molecules our system would still be faster due to the use of light. In small molecule systems it takes some time until the small molecules reach their target in the case of light this happens within milliseconds. Furthermore, with our system it is not necessary to add any kind of activator molecules, which might effect the results, to the cells. This enables researchers to investigate cell activity with and with out the protein in direct comparison, while the only interference is one light puls.</br></br><h2>References</h2></br></br><p><a name=ref55.1>55.1</a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2709356/>Pratt and MacRae (2009) The RNA-induced silencing complex: a versatile gene-silencing machine.</a></p> </br> <p><a name=ref55.2 >55.2</a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3220803/>Davis et al. (2011) Small-molecule control of protein degradation using split adaptors.</a></p></br><p><a name=ref55.3 >55.3</a> <a href=http://www.sciencedirect.com/science/article/pii/S0531556506002798>Bartke (2006) New findings in transgenic, gene knockout and mutant mice.</a></p>";

+

content.type="Background";

+

break;

+

case 102:

+

content.i = 102;

+

content.parents=[101];

+

content.childs=[];

+

content.titleShort = "Student Convention"

+

content.titleLong = "4. BIO.NRW ( PhD ) Student Convention";

+

content.summary= "The iGEM Teams Bonn and Bielefeld were this year represented at the 4th BIO.NRW (PhD) Student Convention in the Esprit-Arena in Düsseldorf. Here our ambassadors Marc and Dustin were able to successfully introduce the current project of light induced protein dregadation in a short presentation. ";

+

content.text= "One hundred undergraduates, graduate students and PhD students from the life sciences came on Friday 12 July 2013 in the Esprit Arena in Dusseldorf, to train with professionals from academia and industry in two days for a successful start of their career. The fourth BIO.NRW ( PhD ) Student Convention, offered an exiting combination of lectures and workshops, regarding non-scientific knowledge and soft skills to complement the expertise of the young researchers. </br></br> We decided to use the opportunity to evaluate the level of knowledge about synthetic biology and iGEM a scientific community. Further, we used our chance to give a presentation and inform the young scientists about synthetic biology, iGEM and our project. In this environment it was a grate experience to discuses our ideas with a scientific community, not only during our presentation but also during the evening event with a barbecue on the sidelines in the Esprit Arena. </br></br> The Presentation of our iGEM project by Dustin Dankelman and Marc Schulte and the iGEM team from Bielefeld started the Convention together with three keynote lectures.The three keynote lectures by representatives of famous companies provided the participants with a practical and intuitive representation of the work of the fields and developments in the industry. Dr. Holger Bengs of the BCNP Consultants GmbH gave the interested participants at the beginning an overview of the biotech industry. In the second lecture, Dr. Barbara Maertens , Head of protein expression of the Cube Biotech GmbH, talked about her jump from a global biotech company to a start-up. The third keynote lecture with the topic research and development in the global pharmaceutical industry was held by Dr. Thomas Lauterbach, Head of Clinical Operations Europe at the biopharmaceutical company UCB. The second day was dedicated to a customized workshop program. </br></br> To evaluate how many scientist know about iGEM, how they think about synthetic biology in general and useful our presentation was, we have designed a questionnaire. At this point we would like to thank all the young scientist, who helped use with their feedback, to address this questions as shown below. <h2>Pictures</h2><div class=subpage-text><table><tr> <th><a href=https://static.igem.org/mediawiki/2013/a/ac/Bonn_MS_BioNRW1.jpg><img src=https://static.igem.org/mediawiki/2013/a/ac/Bonn_MS_BioNRW1.jpg width=260px></a></th> <th><a href=https://static.igem.org/mediawiki/2013/7/7b/Bonn_MS_BioNRW2.jpg><img src=https://static.igem.org/mediawiki/2013/7/7b/Bonn_MS_BioNRW2.jpgwidth=260px></a></th> <th><a href=https://static.igem.org/mediawiki/2013/d/d0/Bonn_MS_BioNRW3.JPG><img src=https://static.igem.org/mediawiki/2013/d/d0/Bonn_MS_BioNRW3.JPG width=260px></a></th></tr><tr> <th><a href=https://static.igem.org/mediawiki/2013/f/f1/Bonn_MS_BioNRW4.JPG><img src=https://static.igem.org/mediawiki/2013/f/f1/Bonn_MS_BioNRW4.JPG width=260px></a></th> <th><a href=https://static.igem.org/mediawiki/2013/1/1d/Bonn_MS_BioNRW5.jpg><img src=https://static.igem.org/mediawiki/2013/1/1d/Bonn_MS_BioNRW5.jpg width=260px></a></th> <th><a href=https://static.igem.org/mediawiki/2013/0/03/Bonn_MS_BioNRW6.JPG><img src=https://static.igem.org/mediawiki/2013/0/03/Bonn_MS_BioNRW6.JPG width=260px></a></th> </tr> </table></br> <h2> Analysis of the questionnaires: </h2></br> 72 people have been asked:</br><img src=https://static.igem.org/mediawiki/2013/6/63/DoYouKnowWhatIgemIs.jpg width=800px></br></br></br><img src=https://static.igem.org/mediawiki/2013/b/b7/HowDoYouRate.jpg width=800px></br>Please klick here to see our <a href=https://static.igem.org/mediawiki/2013/1/16/Fragebogen_iGem.pdf>questionnaire</a>";

+

content.type="Human Practice";

+

break;

@@ Line 266: / Line 266: @@
 content.childs=[35];
 content.titleShort = "pDawn";
-content.titleLong = "";
+content.titleLong = "pDawn system"; (langer Titel)
-content.summary= "";
+content.summary= "The plasmid pDawn was designed by Ohlendorf et al.<sup><a href=#ref21.1>21.1</a></sup> in 2012 together with its counter plasmid pDusk. Both plasmids are single plasmid systems, which allow the activation (pDawn) or repression (pDusk) of gene expression by blue light. They are easy to implement in the laboratory and lead to up to 460-fold activity change upon ilumination.";
-content.text= "";
+content.text= "The plasmid pDawn was designed by Ohlendorf et al.<sup><a href=#ref21.1>21.1</a></sup> in 2012 together with its counter plasmid pDusk. Both plasmids are single plasmid systems, which allow the activation (pDawn) or repression (pDusk) of gene expression by blue light. They described their system as easy to implement in the laboratory and suitable for both preparative and analytical purposes. Further, they were able to show up to 460-fold induction upon ilumination using their system. </br></br> Because, pDawn (and pDusk) is a system that regulates gene expression and thus acts on DNA level it is an eligible candidate for us to compare the speed of action of our system, which acts on protein level to an DNA regulation system. As an additional benefit, with pDawn, we implement a powerful light-induceable system, with low background expression and high dark/light expression difference in the iGEM database. </br></br> Ohlendorf et al. (2012)<sup><a href=#ref21.1>21.1</a></sup> had previously designed the histidine kinase YF1, which phosphorylates FixJ in the absence of blue light. The phosphorylated FixJ drives robust gene expression from the FixK2 promotor as shown in figure 1 for both pDawn and the pDusk. Upon light absorption, net kinase activity of YF1 and consequently gene expression under control of the FixK2 promotor is greatly reduced. pDusk an pDawn enable light-repressed or light-induced gene expression in Escherichia coli and are easy to implement in the laboratory. Therefore, all components of the YF1/FixJ TCS were assembled on only one medium-copy plasmid in which YF1 and FixJ are constitutively expressed by the Lac<sup>q</sup> promotor. Target genes can be introduced via the multiple-cloning site (MCS) under the control of the pFixK2 promotor, allowing light-repressed gene expression. To obtain pDawn the light effect is inverted by the introduction of a gene-inversion cassette into pDusk. This gene-inversion cassette is based on the &lambda; phage repressor cI and the &lambda; phage promotor pR. (see figure 1)</br></br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/a/a1/Bonn_MS_Figure1_pDusk%26pDawn.jpg></br>Figure 1: Plasmids pDusk an pDawn. YF1 and FixJ are constitutively expressed by the Lacq promotor. Target genes can be introduced via the multiple-cloning site (MCS) under the control of the pFixK2 promotor. (Ohlendorf et al. 2012<sup><a href=#ref21.1>21.1</a></sup>)</div></br>Ohlendorf et al. (2012)<sup><a href=#ref21.1>21.1</a></sup> analyzed the system by the insertion of the red-fluorescent reporter protein DsRed Express2. That way they were able to measure up to 460-fold induction upon ilumination. However, they also found that their plasmid, which acts on gene expression level, only allows light-induced perturbations on the timescale of hours.</br></br><h2>References</h2></br><p><a name=ref21.1>21.1</a> <a href=http://www.sciencedirect.com/science/article/pii/S0022283612000113> Ohlendorf et al. (2009) From Dusk till Dawn One-Plasmid Systems for Light-Regulated Gene Expression.</a></p>";
 content.type="Background";
 break;
@@ Line 316: / Line 316: @@
+case 36:
 content.i = 36;
-content.parents=[19];
+content.parents=[37, 19];
-content.childs=[37];
+content.childs=[];
-content.titleShort = "LOV-ipaA - VinD1";
+content.titleShort = "LOV-ipaA & VinD1";
-content.titleLong = "";
+content.titleLong = "LOV-ipaA & VinD1";
-content.summary= "";
+content.summary= "The LOV-ipaA -vinculin system is a combined system for light inducible heterodimerisation. This powerful tool, which allows photocontroled complex formation was establish by Lungu et al. in 2012.";
-content.text= "";
+content.text= "The LOV-ipaA -vinculin system is a combined system for light inducible heterodimerisation. It consists out of a LOV domain, which undergoes conformational changes upon irradiation with blue light, and the ipaA-vinculin hybridization system. This two building blocks have be combined and described by Lungu et al. in 2012.</br></br> Lungu et al. (2008) where able to measure a 49-fold difference in target binding upon irradiation as compared to the dark state. However, they further modified the system by mutations of the LOV-ipaA construct and successfully weakend the baseline affinity for vinculin (initial design: 3.5 nM to 69 nM; mutant: 2.4 nM to >40µM affinity for vinculin) to reduce the dark state activity. </br></br> Lov-ipaA-VinD1 is a powerful tool which allows photocontroled complex formation. To establish this system Lungu et al. (2012)<sup><a href=#ref36.1>36.1</a></sup> fused the Ja helix of the LOV Domain to ipaA.</br>To be more precise they used the LOV2 domain from Avena sativa photopropin 1 (AsLOV2), which – as previously shown – can be used to photomodulate the affinity of peptides for their binding partners (see Figure 1). </br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/0/02/Bonn_MS_Figure1_LOV.jpg></br>Figure 1: General design of AsLOV2 fusion proteins (Lungu et al. 2012)<sup><a href=#ref36.1>36.1</a></sup></div> </br>In other studies had been shown that the LOV domain can be fused to entire protein domains, allowing photomodulation of the protein binding. However, they stated that it might be of high importance to bring the LOV domain closer to ipaA, in order to allow photomodulation in this case, because ipaA is only a peptide and thus more flexible than folded domains.</br></br>Therefore, Lungu et al. (2012)<sup><a href=#ref36.1>36.1</a></sup> identified similar amino acid sequences in the ipaA peptide and the Ja helix of the LOV Domain and used this combined with molecular modeling to create photomodulateable AsLOV2-ipaA (see Figure 2). </br><div class=contant-image><img src=https://static.igem.org/mediawiki/2013/5/54/Bonn_MS_Figure2_LOV-ipaA.jpg></br>Figure 2: Light-inducible LOV-ipaA construct (Lungu et al. 2012)<sup><a href=#ref36.1>36.1</a></sup></div></br>They were able to proof the functionality of the AsLOV2-ipaA system by heterodimerization in yeast (yeast two-hybrid system or Y2H) The yeast two-hybrid system can be used to monitor protein–protein interactions between two proteins. The system is based on a transcription factor, which is split into two separate fragments, called the binding domain (BD) and activating domain (AD). Each domain is fused to one protein and thus only if the proteins interact with each other BD and AD are close enough to initiate the transcription of a reporter gene.</br></br>The basic principle of the LOV-ipaA & VinD1 system works as follows. In the dark state the fusion product LOV-ipaA is not able to form a complex with vinculin, because LOV blocks ipaA sterically. However, activation of the LOV domain with blue light induces conformational changes in the fused molecule, which results in a movement of the Ja helix with the ipaA away from LOV. Thereby, ipaA becomes accessible for VinD1 and a Complex is formed.</br></br>The activation is reversible and the entire system can be genetically encoded. This two facts are the main advantages of this system in contrast to other typically used systems, which like for the chemical system for example, are based on in vivo covalently modified peptides, that can be activated by light induced cleavage. Moreover, the protein used are relatively small and thus should interfere as little as possible with the prokaryotic metabolism, the activity change form dark to light state is high, the system is completely genetically encoded and reversible. But, the most important property of this system is that it allows the light-controlled heterodimerisation of the two split variants of sspB, which is necessary for our system.</br></br><h2>References</h2></br><a name=ref36.1>36.1</a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3334866/>Lungu et al. (2012) Designing photoswitchable peptides using the AsLOV2 domain</a>";
 content.type="Background";
 break;
@@ Line 638: / Line 639: @@
 break;
+case 55:
+content.i = 55;
+content.parents=[53];
+content.childs=[];
+content.titleShort = "Protein function";
+content.titleLong = "Analysis of protein function";
+content.summary= "To understand the role of a specific gene or DNA region is one of the big challenges in lifescience research. Our system, which allows the fast and convenient elimination of defined proteins, is a new improved technique, with many advantages.";
+content.text= "To understand the role of a specific gene or DNA region is one of the big challenges in lifescience research. Our system, which allows the fast and convenient elimination of defined proteins, is a new improved technique, with many advantages. It allows the control of protein activity with respect to time and space, is fast, robust, can be used for different proteins and changes the protein sequence only little, because the ssrA tag consist only out of 15 amino acids. Recently technologies like gene knockout or knockdown had been developed, which allow to investigate the role of a particular gene or DNA region by comparing the knockout organism to a wildtype with a similar genetic background. </br></br> A knockout means that a particular gene is deleted from the genome of an organism. This organism might be bacteria or yeast, but also eukaryotic cells, plants or even animals. To create a knockout organism recombinant DNA is inserted into a gene (Bartke, 2006<sup><a href=#ref55.3>55.3</a></sup>). When a genes sequence is interrupted, it may still be translated, but the resulting protein will be nonfunctional. Moreover, it is possible to knockout the gene only in defined tissues or at defined time points. This technique is called a conditional knockout. </br></br> On the other hand the knockdown, does not eliminate the specific gene on DNA, but on RNA level. Here interfering RNAs (siRNA) are inserted into the cell, leading to the degradation of the genes mRNA (Pratt and MacRae, 2009<sup><a href=#ref55.1>55.1</a></sup>) and hence no protein can be produced. </br></br> In contrast to the knockout and knockdown, our system allows the expression of the gene and the translation into functional protein. However, irradiation with blue light leads to the fast elimination of the particular protein. Therefore, one of the major advantages of our system is its speed, not only in comparison to knockout and knockdown, but also in comparison to other protein level systems. Comparing our system, to other protein level systems, like the system developed by Davis et al.<sup><a href=#ref55.2>55.2</a></sup> in 2011, which are induced by small molecules our system would still be faster due to the use of light. In small molecule systems it takes some time until the small molecules reach their target in the case of light this happens within milliseconds. Furthermore, with our system it is not necessary to add any kind of activator molecules, which might effect the results, to the cells. This enables researchers to investigate cell activity with and with out the protein in direct comparison, while the only interference is one light puls.</br></br><h2>References</h2></br></br><p><a name=ref55.1>55.1</a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2709356/>Pratt and MacRae (2009) The RNA-induced silencing complex: a versatile gene-silencing machine.</a></p> </br> <p><a name=ref55.2 >55.2</a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3220803/>Davis et al. (2011) Small-molecule control of protein degradation using split adaptors.</a></p></br><p><a name=ref55.3 >55.3</a> <a href=http://www.sciencedirect.com/science/article/pii/S0531556506002798>Bartke (2006) New findings in transgenic, gene knockout and mutant mice.</a></p>";
+content.type="Background";
+break;
+case 102:
+content.i = 102;
+content.parents=[101];
+content.childs=[];
+content.titleShort = "Student Convention"
+content.titleLong = "4. BIO.NRW ( PhD ) Student Convention";
+content.summary= "The iGEM Teams Bonn and Bielefeld were this year represented at the 4th BIO.NRW (PhD) Student Convention in the Esprit-Arena in Düsseldorf. Here our ambassadors Marc and Dustin were able to successfully introduce the current project of light induced protein dregadation in a short presentation. ";
+content.text= "One hundred undergraduates, graduate students and PhD students from the life sciences came on Friday 12 July 2013 in the Esprit Arena in Dusseldorf, to train with professionals from academia and industry in two days for a successful start of their career. The fourth BIO.NRW ( PhD ) Student Convention, offered an exiting combination of lectures and workshops, regarding non-scientific knowledge and soft skills to complement the expertise of the young researchers. </br></br> We decided to use the opportunity to evaluate the level of knowledge about synthetic biology and iGEM a scientific community. Further, we used our chance to give a presentation and inform the young scientists about synthetic biology, iGEM and our project. In this environment it was a grate experience to discuses our ideas with a scientific community, not only during our presentation but also during the evening event with a barbecue on the sidelines in the Esprit Arena. </br></br> The Presentation of our iGEM project by Dustin Dankelman and Marc Schulte and the iGEM team from Bielefeld started the Convention together with three keynote lectures.The three keynote lectures by representatives of famous companies provided the participants with a practical and intuitive representation of the work of the fields and developments in the industry. Dr. Holger Bengs of the BCNP Consultants GmbH gave the interested participants at the beginning an overview of the biotech industry. In the second lecture, Dr. Barbara Maertens , Head of protein expression of the Cube Biotech GmbH, talked about her jump from a global biotech company to a start-up. The third keynote lecture with the topic research and development in the global pharmaceutical industry was held by Dr. Thomas Lauterbach, Head of Clinical Operations Europe at the biopharmaceutical company UCB. The second day was dedicated to a customized workshop program. </br></br> To evaluate how many scientist know about iGEM, how they think about synthetic biology in general and useful our presentation was, we have designed a questionnaire. At this point we would like to thank all the young scientist, who helped use with their feedback, to address this questions as shown below. <h2>Pictures</h2><div class=subpage-text><table><tr>            <th><a href=https://static.igem.org/mediawiki/2013/a/ac/Bonn_MS_BioNRW1.jpg><img src=https://static.igem.org/mediawiki/2013/a/ac/Bonn_MS_BioNRW1.jpg width=260px></a></th>            <th><a href=https://static.igem.org/mediawiki/2013/7/7b/Bonn_MS_BioNRW2.jpg><img src=https://static.igem.org/mediawiki/2013/7/7b/Bonn_MS_BioNRW2.jpgwidth=260px></a></th>            <th><a href=https://static.igem.org/mediawiki/2013/d/d0/Bonn_MS_BioNRW3.JPG><img src=https://static.igem.org/mediawiki/2013/d/d0/Bonn_MS_BioNRW3.JPG width=260px></a></th></tr><tr>            <th><a href=https://static.igem.org/mediawiki/2013/f/f1/Bonn_MS_BioNRW4.JPG><img src=https://static.igem.org/mediawiki/2013/f/f1/Bonn_MS_BioNRW4.JPG width=260px></a></th>            <th><a href=https://static.igem.org/mediawiki/2013/1/1d/Bonn_MS_BioNRW5.jpg><img src=https://static.igem.org/mediawiki/2013/1/1d/Bonn_MS_BioNRW5.jpg width=260px></a></th>            <th><a href=https://static.igem.org/mediawiki/2013/0/03/Bonn_MS_BioNRW6.JPG><img src=https://static.igem.org/mediawiki/2013/0/03/Bonn_MS_BioNRW6.JPG width=260px></a></th>           </tr>          </table></br>            <h2> Analysis of the questionnaires: </h2></br>             72 people have been asked:</br><img src=https://static.igem.org/mediawiki/2013/6/63/DoYouKnowWhatIgemIs.jpg width=800px></br></br></br><img src=https://static.igem.org/mediawiki/2013/b/b7/HowDoYouRate.jpg width=800px></br>Please klick here to see our <a href=https://static.igem.org/mediawiki/2013/1/16/Fragebogen_iGem.pdf>questionnaire</a>";
+content.type="Human Practice";
+break;

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Revision as of 21:05, 1 October 2013