Team:Freiburg/Project/toolkit

You have chosen to do a gene activation experiment using a non inducible Cas9-VP16 device. Therefor you first have to order the following plasmids from the iGEM parts registry :

Note:
All plasmids are optimised for an expression in mammalian systems. The devices are also available containing an SV40 instead of an CMV promotor (have a look at our parts side). This system was tested mainly in CHO, HEK-293-T and HeLa cells.

Experimental design (recommendation for a mammalian cell culture):

1. Transfect the appropriate crRNA - plasmid togehter with BBa_K1150017 that has no effector for control
2. Transfect BBa_K1150020 without any crRNA plasmid as off target control
3. Transfect BBa_K1150020 with all desired crRNA plasmids (seperate and in combinitions)