Our parts

We managed to submit 4 new parts that will give other teams the possibility to reproduce our experiments with both the switches we tested.

The melanopsin coding gene and the NFAT response element will allow to use the more complex switch, by just cloning melanopsin into a mammalian vector and clone a readout gene (fluorescent protein, quantitatively measurable protein) downstream NFAT and clone both in a mammalian vector.

We also submitted our favorite parts, which are a mammalian LovTAP-VP16 and a composite readout system build to respond to it. Both parts only need cloning into a mammalian vector to be ready for transfection.

<groupparts>iGEM012 EPF-Lausanne</groupparts>

How to use

1) Cloning:

First clone the part into a mammalian expression vector such as pcDNA3.1(+)[http://products.invitrogen.com/ivgn/product/V79020?ICID=search-product] or pCEP4[http://products.invitrogen.com/ivgn/product/V04450]. These are the two main expression vectors we used. To use the melanopsin pathway, you also need to clone a target gene of your choice downstream pNFAT.

• ->

2) Choose your switch:

• Simple switch = LovTAP-VP16 + LovTAP readout

• Complex switch = Melanopsin + pNFAT-target gene

3) Transfection Co-transfect the combination of photoreceptive protein and readout construct of your choice in mammalian cells!