The 2016 BostonU iGEM team was a nominee for the Best Basic Part Award.
To create the Gemini parts library, we made use of two “classes” of plasmids: guide RNA expressing vectors and guide RNA operator reporter vectors. When these two plasmids are co-transfected with a dCas9-VPR plasmid into a mammalian cell line, a gene of interest is expressed. Our work for iGEM dealt with fluorescent proteins; specifically, GFP for the parts submitted to the registry.
The guide RNA expressing vector can be considered a basic part. Its design consists of a hU6 promoter upstream of a guide RNA sequence, which has a target recognition sequence and a scaffold sequence for dCas9 binding, followed by a polyT terminator. This basic part, whose data can be found on BBa_K1875012, was based off of the work by Feng Zhang at MIT and the x330 plasmid. Our part here expresses gRNA 8 (g8) from the Gemini parts library. It represents what we feel is one of best basic parts in our library due to its consistency in experimentation.