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Team:Freiburg/Project/Robot
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<br><div align="justify">Today, only two types of TAL effectors exist: TAL transcription factors and TALENS. We believe that many more effectors can be fused to the TAL protein. We therefore created a platform for future iGEM students that allows them to easily produce their own new classes of TAL effectors. We are actually still working on two of them (but have no results yet): We have fused the catalytic domain of Suv 39 H1 to the c-terminus of our TAL scaffold to set histone marks in a sequence specific manner. Unfortunately, optimizing a chip assay (which, in combination with qPCR is our readout for this effector) takes much longer than expected, but we hope, that we will one day be able to open the field of epigenetics to synthetic biology. Another interesting project we have started working on are TAL effector recombinases. These n-terminal fusion proteins of TALEs and serine recombinases. Interesting about serine recombinases is that their DNA binding domain is spatially distinct from their catalytic domain. We have started to replace the natural DNA binding domain by our TAL effector in order to induce recombination at any point within a genome. This, obviously, would be the holy grail for synthetic biology. We believe that the potential for new types of TAL effectors is huge and we are looking forward to seeing new such technologies arise from future iGEM competitions. | <br><div align="justify">Today, only two types of TAL effectors exist: TAL transcription factors and TALENS. We believe that many more effectors can be fused to the TAL protein. We therefore created a platform for future iGEM students that allows them to easily produce their own new classes of TAL effectors. We are actually still working on two of them (but have no results yet): We have fused the catalytic domain of Suv 39 H1 to the c-terminus of our TAL scaffold to set histone marks in a sequence specific manner. Unfortunately, optimizing a chip assay (which, in combination with qPCR is our readout for this effector) takes much longer than expected, but we hope, that we will one day be able to open the field of epigenetics to synthetic biology. Another interesting project we have started working on are TAL effector recombinases. These n-terminal fusion proteins of TALEs and serine recombinases. Interesting about serine recombinases is that their DNA binding domain is spatially distinct from their catalytic domain. We have started to replace the natural DNA binding domain by our TAL effector in order to induce recombination at any point within a genome. This, obviously, would be the holy grail for synthetic biology. We believe that the potential for new types of TAL effectors is huge and we are looking forward to seeing new such technologies arise from future iGEM competitions. | ||
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Revision as of 19:54, 24 October 2012
Automating TAL production
Because of the simplicity of our toolkit, it is easily possible to automate TAL production. This will help to meet the high demand for TAL effectors and at the same time further reduce costs of TAL production.
How it is today
Ordering TAL effectors today can be a tough decision for a scientist. Not only is it complicated but also unbelievably expensive and time consuming. We checked some of the commercially available TAL products and found prices up to 5000 Euros for production of just one custom TAL nuclease pair. For support, it was necessary to pay another 1000 Euros and you had to wait for several weeks until your TAL was finished and shipped.
How it will be tomorrow
The market for TAL effectors, especially TAL nucleases, is dominated by only a few companies holding patents on currently available TAL production methods. These companies are setting prices for TAL effectors and make a living selling overpriced products to scientist all over the world.
With our toolkit this practice hopefully comes to an end, as we shift the production of high quality TAL proteins into your hands. Our toolkit is not only easy to use and affordable, it is also automatable. The few steps that are necessary to produce TAL proteins with our kit are easily done by a pipetting robot in virtually no time. You can have 96 different TAL effectors on one plate in one run, put all of them into a thermocycler and use them directly afterwards.
Automated TAL production with a pippeting robot
Inventing new classes of TALEs
Today, only two types of TAL effectors exist: TAL transcription factors and TALENS. We believe that many more effectors can be fused to the TAL protein. We therefore created a platform for future iGEM students that allows them to easily produce their own new classes of TAL effectors. We are actually still working on two of them (but have no results yet): We have fused the catalytic domain of Suv 39 H1 to the c-terminus of our TAL scaffold to set histone marks in a sequence specific manner. Unfortunately, optimizing a chip assay (which, in combination with qPCR is our readout for this effector) takes much longer than expected, but we hope, that we will one day be able to open the field of epigenetics to synthetic biology. Another interesting project we have started working on are TAL effector recombinases. These n-terminal fusion proteins of TALEs and serine recombinases. Interesting about serine recombinases is that their DNA binding domain is spatially distinct from their catalytic domain. We have started to replace the natural DNA binding domain by our TAL effector in order to induce recombination at any point within a genome. This, obviously, would be the holy grail for synthetic biology. We believe that the potential for new types of TAL effectors is huge and we are looking forward to seeing new such technologies arise from future iGEM competitions.
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