Difference between revisions of "Team:NAU-CHINA/testh"

 
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Fig4. Function verification of TEV elimination of modified tetR inhibition with eukaryotic cells
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Overview
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Stably transfer Jurkat T cells with the modified TetR gene to construct a stably transferred cell line. Then transfer plasmids containing Lag16 - synNotch-TEV and tetO-miniCMV-EGFP genes into the aforementioned stably transferred cell. Co-culture the 293T cells expressing GFP on the cell surface with these Jurkat T cells for 4 h when 293T cells were deposited at the bottom of the culture medium and separated from suspended Jurkat T cells.  
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We developed iGEMCloud to solve some of iGEM's current problems, with its immediacy, openness and interactivity. At present, all the information about iGEM comes from many websites. They are scattered, cluttered and unfriendly to personal user.
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(A) Fluorescence microscope observation of the stably transfferred cell line.  
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(B) Fluorescence microscope observation of the stably transfferred cell line transferred plasmids containing Lag16 - synNotch-TEV and tetO-miniCMV-EGFP genes.
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So we have created this application(So far, it is android based, and we hope to launch the iOS version in the future) , which is hoped to be a information gathering platform that receives the latest information of iGEM, a micro-blog that allows all users to participate in discussions in group or individual, and a community that built for iGEMers all around the world.
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(C) Fluorescence microscope observation of the aforementioned Jurkat T cells co-cultured with 293T cells expressing cell surface-expressed GFP for 4 h.
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Through fluorescence microscopy, we could observe that the suspended T cells emit green fluorescence, which is clearly distinguished from the weaker green fluorescence of 293T cells expressing surface-expressed GFP  deposited at the bottom of the culture medium. The results show that TEV can relieve the inhibition of tetR on the promoter in 293T cells. It means that we have successfully verified the function of TEV - activated transcription system based on the modified tetR in eukaryotic cells and the results also confirm preliminarily that our upstream circuit can work normally. However, we have to admit that due to we chosed GFP as our reporter gene, it is difficult to distinguish it from cell surface-expressed GFP. Our verification experiment is not intuitive. If we need to prove the function of TEV suppressing the inhibition of tetR strongly, further  optimized experiments are still needed.
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Instructions
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Downstream circuit
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1.Register an account with your email address, enter the verify code, and use this address to log in.
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The downstream pathway is the core circuit for us to realize the threshold function. According to literature  [3], they have verified the inversion function of the three recombinases in prokaryotic cells and proved the threshold function of the recombinases, i.e. the recombinases do not have the inversion function at low concentration. Only when the concentration of recombinase reaches a certain threshold, can the recombinases work normally.
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According to the same document, we designed our pathway in eukaryotic cells, expecting to realize threshold switching in eukaryotic cells. For this reason, we try to test the inversion function of recombinases and the threshold characteristics of the combination of three different recombinases ( Bxb1, TP901, φ C31 ) and three promoters with different intensities ( miniCMV, EF1 - α, Ubc ) in eukaryotic cells.
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2.In the application, you can receive official news, notifications, etc. And so that you can get the latest news of iGEM events in a timely, convenient and accurate way.
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We also verify the function of RDF [4] to demonstrate our 0/1 switch resettable in HEK 293T cells.
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3.Users can write blog posts in the discussion forum for all users to browse. The blog post is in the form of pictures + text. The blog posts are classified into different sections, such as experiment, seek help, etc. And so that you can share your ideas with all iGEM participants.
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Excellent blog posts will be tagged so that they are discovered by users. At the same time, users who see the blog posts written by you can comment or reply to you. The content of the reply is in the form of image + text. When you find a nice blog post, click on the star symbol in the upper right to bookmark it for later reading.
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4.If you want to remind specific users to view the content you have posted, you can remind the user to view this blog post or comment by "@" + "user nickname".
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5.You can search for blog posts in one or more keywords in the app, and sort the search results by time, number of view , and collection. You can also search for users or teams.
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6.You can follow the teams or users that you think are worthy of attention so that you can view their published content more easily in the future.
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7.Each user can create or join a team, become a member, and use their team space. The team creator is the captain who can edit the team information in the team space for others to join and show the team research direction. Only members of the team can post in the team space, but everyone can view it. Also, the captain can add albums in the team space,where members of the team can post photos.
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8.This app has built-in gadgets that may be needed for the lab and more other features will be added in the future...
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What it could be
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An information gathering platform:
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We will gather information from other website to release, and we hope to build contact with some organization to get instant and accurate information. The user of this app can share their information in it. If it is valuable and reliable, we will push it to everyone.
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A platform for interaction between teams:
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Through the form of the team space, each team can easily interact with other teams. This will strengthen the connection between iGEM teams and cause countless positive possibilities.
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 +
An iGEM community:
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Each iGEMer could leave a footprint here and show themselves here. Everyone here is part of the iGEM.
 +
 
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The App we conceived can provide comprehensive information on the release of events, each team to register their own team home page, various activities related to iGEM, question answer and experience sharing , topic discussion, experimental supplies, and more.

Latest revision as of 10:05, 17 October 2018

Overview We developed iGEMCloud to solve some of iGEM's current problems, with its immediacy, openness and interactivity. At present, all the information about iGEM comes from many websites. They are scattered, cluttered and unfriendly to personal user.

So we have created this application(So far, it is android based, and we hope to launch the iOS version in the future) , which is hoped to be a information gathering platform that receives the latest information of iGEM, a micro-blog that allows all users to participate in discussions in group or individual, and a community that built for iGEMers all around the world.

Instructions 1.Register an account with your email address, enter the verify code, and use this address to log in.

2.In the application, you can receive official news, notifications, etc. And so that you can get the latest news of iGEM events in a timely, convenient and accurate way.

3.Users can write blog posts in the discussion forum for all users to browse. The blog post is in the form of pictures + text. The blog posts are classified into different sections, such as experiment, seek help, etc. And so that you can share your ideas with all iGEM participants.

Excellent blog posts will be tagged so that they are discovered by users. At the same time, users who see the blog posts written by you can comment or reply to you. The content of the reply is in the form of image + text. When you find a nice blog post, click on the star symbol in the upper right to bookmark it for later reading.

4.If you want to remind specific users to view the content you have posted, you can remind the user to view this blog post or comment by "@" + "user nickname".

5.You can search for blog posts in one or more keywords in the app, and sort the search results by time, number of view , and collection. You can also search for users or teams.

6.You can follow the teams or users that you think are worthy of attention so that you can view their published content more easily in the future.

7.Each user can create or join a team, become a member, and use their team space. The team creator is the captain who can edit the team information in the team space for others to join and show the team research direction. Only members of the team can post in the team space, but everyone can view it. Also, the captain can add albums in the team space,where members of the team can post photos.

8.This app has built-in gadgets that may be needed for the lab and more other features will be added in the future...

What it could be An information gathering platform: We will gather information from other website to release, and we hope to build contact with some organization to get instant and accurate information. The user of this app can share their information in it. If it is valuable and reliable, we will push it to everyone.

A platform for interaction between teams: Through the form of the team space, each team can easily interact with other teams. This will strengthen the connection between iGEM teams and cause countless positive possibilities.

An iGEM community: Each iGEMer could leave a footprint here and show themselves here. Everyone here is part of the iGEM.

The App we conceived can provide comprehensive information on the release of events, each team to register their own team home page, various activities related to iGEM, question answer and experience sharing , topic discussion, experimental supplies, and more.