Team:BostonU/Part Collection
The BostonU 2016 iGEM team created Gemini, a design space that combines digital and analog expression systems to easily modulate exogenous gene expression levels in human cells. The system relies on three components: a genome-orthogonal gRNA recognizes a corresponding DNA operator sequence upstream of a minimal promoter, and it recruits dCas9-VPR to transactivate the output gene. We developed a set of mutually-orthogonal gRNAs to enable multiplexed gene regulation without cross-talk, and a set of gRNA-operator plasmids to achieve varied expression levels. Our submitted parts collection has 2 gRNA expression devices (BBa_K1875011-BBa_K1875012) and 7 gRNA-operator devices (BBa_K1875013-BBa_K1875019). We validated these parts using flow cytometry. We demonstrated “digital” expression when comparing output gene activation with or without gRNAs, and “analog” expression when comparing different gRNA-operator architectures (single, multimerized, and mutated sequences).
The team created pages for parts BBa_K1875000 - BBa_K1875019 and submitted parts BBa_K1875011 - BBa_K1875019.
