Toggle navigation Home Team Team Media Collaborations Sponsors Acknowledgements Project Background Design CRISPR/Cas9 Strategy Experiments Notebook Results Perspective Interlab Study Parts Parts Basic Parts Composite Parts Human Pratices Overview Societal Issues of CRISPR/Cas9 Responsible Research and Innovation GMO regulation Integrated Practices Engagement Model Attributions Safety Contents 1 Project description 1.1 Welcome to iGEM 2016! 1.1.1 Before you start: 1.1.2 Styling your wiki 1.1.3 Wiki template information 1.1.4 Editing your wiki 1.1.5 Tips 1.1.6 Inspiration 1.1.7 Uploading pictures and files Project description Recent observations lead to the idea that genes, not in the same operon but spatially close, are highly co-transcribed, even in the absence of regulatory factors at their promoter regions. The iGEM Paris-Saclay project aims to study the effects of DNA topology on gene expression in E.coli by answering to this question: Does bringing a strong promoter closer to a weak promoter influence the expression level of genes located downstream? We have designed a new tool based on CRISPR/Cas9 system to bring two specific DNA regions closer. This system is composed of two different dCas9 fused with each part of FRB / FKBP12 dimerization system. Each dCas9 will target a specific DNA sequence, one on the chromosome and one on a plasmid, whereas dimerization system will promote the joining of the two dCas9 when rapalog is added. In order to assess whether or not this system works, we have also designed a new tool to visualize the interaction between both dCas9. This tool is composed of a split GFP attached to two dCas9. These two small GFP tags will interact with the complementary GFP detector only if the two dCas9 are close enough to interact. If we obtain a higher expression level of the weak promoter with our two tools, it could lead to several useful applications. For example, we would be able to use this tool to enhance gene expression of any endogenous genes due to CRISPR/Cas9 specificity. Indeed, it would be possible to design specific sgRNA but the user should be aware about off-target activity of the CRISPR/Cas9 system. A MODIFIER CAR MÊME DESCRIPTION QUE L'OVERVIEW DU PROJET A COMPLÉTER AVEC LA VIDEO D'ALICE Welcome to iGEM 2016! Your team has been approved and you are ready to start the iGEM season! Before you start: Please read the following pages: <a href="https://2016.igem.org/Requirements">Requirements page </a> <a href="https://2016.igem.org/Wiki_How-To">Wiki Requirements page</a> <a href="https://2016.igem.org/Resources/Template_Documentation"> Template Documentation </a> Styling your wiki You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project. While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided. Wiki template information We have created these wiki template pages to help you get started and to help you think about how your team will be evaluated. You can find a list of all the pages tied to awards here at the <a href="https://2016.igem.org/Judging/Pages_for_Awards/Instructions">Pages for awards</a> link. You must edit these pages to be evaluated for medals and awards, but ultimately the design, layout, style and all other elements of your team wiki is up to you! Editing your wiki On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world! <a href="https://2016.igem.org/wiki/index.php?title=Team:Example&action=edit"> </a>Use WikiTools - Edit in the black menu bar to edit this page Tips This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: State your accomplishments! Tell people what you have achieved from the start. Be clear about what you are doing and how you plan to do this. You have a global audience! Consider the different backgrounds that your users come from. Make sure information is easy to find; nothing should be more than 3 clicks away. Avoid using very small fonts and low contrast colors; information should be easy to read. Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="https://2016.igem.org/Calendar">iGEM 2016 calendar</a> Have lots of fun! Inspiration You can also view other team wikis for inspiration! Here are some examples: <a href="https://2014.igem.org/Team:SDU-Denmark/"> 2014 SDU Denmark </a> <a href="https://2014.igem.org/Team:Aalto-Helsinki">2014 Aalto-Helsinki</a> <a href="https://2014.igem.org/Team:LMU-Munich">2014 LMU-Munich</a> <a href="https://2014.igem.org/Team:Michigan"> 2014 Michigan</a> <a href="https://2014.igem.org/Team:ITESM-Guadalajara">2014 ITESM-Guadalajara </a> <a href="https://2014.igem.org/Team:SCU-China"> 2014 SCU-China </a> Uploading pictures and files You can upload your pictures and files to the iGEM 2016 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name. When you upload, set the "Destination Filename" to T--YourOfficialTeamName--NameOfFile.jpg. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!) UPLOAD FILES </html>
Recent observations lead to the idea that genes, not in the same operon but spatially close, are highly co-transcribed, even in the absence of regulatory factors at their promoter regions.
The iGEM Paris-Saclay project aims to study the effects of DNA topology on gene expression in E.coli by answering to this question: Does bringing a strong promoter closer to a weak promoter influence the expression level of genes located downstream?
We have designed a new tool based on CRISPR/Cas9 system to bring two specific DNA regions closer. This system is composed of two different dCas9 fused with each part of FRB / FKBP12 dimerization system. Each dCas9 will target a specific DNA sequence, one on the chromosome and one on a plasmid, whereas dimerization system will promote the joining of the two dCas9 when rapalog is added. In order to assess whether or not this system works, we have also designed a new tool to visualize the interaction between both dCas9. This tool is composed of a split GFP attached to two dCas9. These two small GFP tags will interact with the complementary GFP detector only if the two dCas9 are close enough to interact.
If we obtain a higher expression level of the weak promoter with our two tools, it could lead to several useful applications. For example, we would be able to use this tool to enhance gene expression of any endogenous genes due to CRISPR/Cas9 specificity. Indeed, it would be possible to design specific sgRNA but the user should be aware about off-target activity of the CRISPR/Cas9 system.
A MODIFIER CAR MÊME DESCRIPTION QUE L'OVERVIEW DU PROJET A COMPLÉTER AVEC LA VIDEO D'ALICE
Your team has been approved and you are ready to start the iGEM season!
Please read the following pages:
You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.
While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.
We have created these wiki template pages to help you get started and to help you think about how your team will be evaluated. You can find a list of all the pages tied to awards here at the <a href="https://2016.igem.org/Judging/Pages_for_Awards/Instructions">Pages for awards</a> link. You must edit these pages to be evaluated for medals and awards, but ultimately the design, layout, style and all other elements of your team wiki is up to you!
On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world!
<a href="https://2016.igem.org/wiki/index.php?title=Team:Example&action=edit"> </a>Use WikiTools - Edit in the black menu bar to edit this page
This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started:
You can also view other team wikis for inspiration! Here are some examples:
You can upload your pictures and files to the iGEM 2016 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name. When you upload, set the "Destination Filename" to T--YourOfficialTeamName--NameOfFile.jpg. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)
T--YourOfficialTeamName--NameOfFile.jpg
UPLOAD FILES
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