Difference between revisions of "Team:MIT/Mammalian synthetic biology kit"

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<p>Our kit is meant to be used with Gateway cloning reactions.  Gateway is a recombinase-based cloning method used by the phage λ against e.Coli, and isolated and optimized for for synthetic biology by Invitrogen.   
 
<p>Our kit is meant to be used with Gateway cloning reactions.  Gateway is a recombinase-based cloning method used by the phage λ against e.Coli, and isolated and optimized for for synthetic biology by Invitrogen.   
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<a href="https://2016.igem.org/Team:MIT/pdestmcherry"><center><img src="https://static.igem.org/mediawiki/2016/7/73/T--MIT--pdestmcherryplasmid.png" style="width:653px;height:518px;"></center></a>
 
<a href="https://2016.igem.org/Team:MIT/pdestmcherry"><center><img src="https://static.igem.org/mediawiki/2016/7/73/T--MIT--pdestmcherryplasmid.png" style="width:653px;height:518px;"></center></a>
 
<p> needs to be linked to the protocol/parts page, but right now linked to what may be a throwaway </p>
 
<p> needs to be linked to the protocol/parts page, but right now linked to what may be a throwaway </p>
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<p>hef1a is a constitutive promoter.  Connected to various fluorescence, these constructs can be used for transfection markers </p>
 
<p>hef1a is a constitutive promoter.  Connected to various fluorescence, these constructs can be used for transfection markers </p>
 
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Revision as of 17:05, 9 October 2016

With the development of efficacious gene editing tools and improved practices, mammalian synthetic biology grows more prevalent. While our project is one example of how synthetic biology can be used to diagnose diseases, we hope that more people are able to build off our work, and the work of others to apply synthetic biology to mammalian cells.

Hoping to see what amazing work can come from future teams and scientists, we have put together a toolbox of parts to make it easier to start mammalian work.

Our kit is meant to be used with Gateway cloning reactions. Gateway is a recombinase-based cloning method used by the phage λ against e.Coli, and isolated and optimized for for synthetic biology by Invitrogen.

There are two plasmids involved:

  • pENTR: the entry plasmids contain the desired genes, flanked by L1 and L2 sites.
  • pDEST: the destination plasmid will eventually be transformed into the E.Coli. There is a selection or screening tool flanked by R1 and R2 sites.

    • From recombination of the L1, L2, R1, and R2 sites, the pEXPR expression plasmid is formed.

    currently, the screening tool used for the pDEST is ccdb. However, a patent on it makes it impossible to distribute to iGEM teams. To allow distribution, we have created a pDEST plasmid that has mCherry fluorescence as a screening tool.

    Along with our pDEST, we include several pENTR vectors, with promoters and useful genes that can be combined and put into eColi to grow up before transfection into mammalian systems

    needs to be linked to the protocol/parts page, but right now linked to what may be a throwaway

    hef1a is a constitutive promoter. Connected to various fluorescence, these constructs can be used for transfection markers