Revision as of 11:25, 28 August 2014

Cambridge iGEM 2014

Project Description

Sensing is an essential aspect of engineering. We need information about the world to make intelligent efforts to manipulate it. Ideal sensors are reliable, accurate and unobtrusive. Plants as biosensors fulfill these requirements and have the further benefit of being inexpensive and self-reproducing.

Our aim is to use the lower plant Marchantia polymorpha as a flexible, open source biosensor. The input, processing and output abilities are each parceled into separate modules which are linked using transcription factors and inducible promoters. Modules can be interchanged, so by swapping one input or output for another, PhytoSpy can flexibly be used to sense any condition or chemical, and report it in any way that a module has been designed for. This modular framework allows many devices to be constructed from the same library of components.

Our dream is to produce input, processing and output modules in separate plants which can be combined through Mendelian crossing. We want to make plant biosensors accessible to the home enthusiast in the same way that electronics has been made accessible by Arduino.

Constructs

35S - Chromoproteins
It seems we've managed to build the following sequences correctly...
eforRed
tsPurple
tsPurple-N7
asPink-N7
amilCP-N7
Now to be transformed into Marchantia.

35S - HAP1 - HAP1 UAS - Chromoproteins
Constructs and primers designed, primers to be checked and ordered.

35S - HAP1 - HAP1 UAS - Raspberry ketone
Benzalacetone synthase: sequence and sample from Japanese team
Benzalacetone reductase: primers designed from the Stanford biobrick, waiting for extraction kits to extract from raspberry plant.

Venus - HHR+Apt

35S - HAP1 - HHR+Apt - HAP1 UAS - Venus
Samples for two buffers, an inverter and a control sent by C. Smolke in a plate.
Primers to order today (04 Aug)
We now have theophylline and tetracycline.

Inducible promoter - GAL4 - GAL4 UAS - HAP1 - HAP1 UAS - Venus
List of nitrate, phosphate inducible promoters, clock genes, light related genes etc. and sequences of corrensponding proteins made. Comparison with tblastn to the marchantia genes done, now analysing the data to retain best candidates and get promoter sequences.

Dexamethasone - GAL4 - GAL4 UAS - HAP1 - HAP1 UAS - Venus
Full cycle - Salil says that we are not doing any Dexamethasone inducible constructs

Auxin inducible promoter - VenusN7
Hugh - initial research: I am trying to find a sample which contains the promoter

Project Description and History

Content

References

Overall project summary
Project Details
Materials and Methods
The Experiments
Results
Data analysis
Conclusions

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 <li>Conclusions</li>
 </ol>
-<h1>Brainstorming Ideas: Imagination on the loose!</h1>
-Many weird and wonderful ideas came up from Radio Plants to Self-reproducible diagnostic tests.<br>
-After some heated debates, dead alley ways, some highs and grief we narrowed down to some final ideas.<br>
-Here, for your amusement, is some of what iGEMCambridge 2014 could have been: <br>
-<h4>Mar-Cam-tia, your Desktop Lamp</h4>
-<p>Engineer the luciferin-luciferase system that the 2010 Cambridge team put into 'E. glowli' into Marchy, and show the dodgy kickstarter dude how it should really be done. This could be linked to the plant's circadian rhythm to ensure only nigh time glowing.</p>
-<h4> Mar-Cam-tia, the volatile factory with some twists</h4>
-<p>Add metabolic pathways that produce perfumed volatiles (eg geraniol, limonene, geosmin). Or insect repellents/attractants such as E-beta-farnesene to repel aphids and attract ladybirds, or bombykol to attract moths from miles away.</p>
-<h4> Mar-Cam-tia, the Night Catcher </h4>
-<p>Production could be linked to circadian clock to change smell (or species of moth attracted) throughout the day -<strong> ie Mar-Cam-tia as a clock.</strong> There could be a link to Marchy's air pores.</p>
-<h4>Mar-Cam-tia, the ultimate multipurpose phytoELISA platform</h4>
-<p>An antigen receptor based on an antibody would be expressed on the surface of Marchantia, and a potentially novel signalling pathway (perhaps involving cAMP production or a tyrosine kinase dimer formation mechanism) engineered such that when the antigen of interest binds with the receptor, downstream events induce an obvious reporting process such as chromoprotein expression, luciferin-mediated glowing, morphological changes or the release of a characteristic smell. Qualitative disease testing with a product that replicates itself out of water and sunshine instead of a single use ELISA that costs $300! Bring on the Start Up!</p>
-<h4>Mar-Cam-tia, a new eco-friendly touch screen</h4>
-<p>If chromoprotein expression can be triggered in arbitrary cells by irradiating them with a far-red laser to induce the phytochrome signalling pathway in them, then thalli could be written on with light and an image formed. Different lasers, different colour chromoproteins = full palette of laser-plant-pens. No paper waste</p>
-<h4>Marchantia, change-o-morpha</h4>
-<p>A morphological study of Marchy: Can we engineer shape? Can we disrupt the normal course of growth and shape formation? Maybe we can shape it like a cauliflower.</p>
-<h4>Taming the Beast</h4>
-<p>Sporlulation is a bio-containment issue. If Marchy was less sexually prolific, we would have less of a problem. It would also be interesting and aesthetically pleasing to have sex specific expression of colour. (quite some controversy about colour selection)</p>
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Team:Cambridge-JIC/Project

From 2014.igem.org