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Overview
In nature, organisms’ genes are robust to the point that the organism persistently reproduces under different external and internal conditions. Synthetic robustness within the field of synthetic biology is a hurdle yet to be crossed. Taguchi method is introduced to reduce variation in gene circuits through robust design of experiment. Three promoters combined with three ribosomal binding sites and three terminators are combined in a series of 27 different circuits each consisting of a promoter followed by an RBS followed by GFP and finished with a terminator. Taguchi method uses orthogonal arrays to organize the parameters and tests for pairs of combinations of factors to gather data with minimum experiments in order to reduce cost and time. The particular gene of interest is GFP, thus, expression level of fluorescence is used as criteria to predict the most robust combination with least variance.Background
One of the overwhelming problems that synthetic biologists face on a daily basis is the variability in genetic constructs. With our ever persistent drive to find a way to develop a standard set of parts for synthetic biology, it is often overlooked that these “standard” parts do not interact the same way that a nut and a bolt do. Sure the threads of each must match for compatibility, but you are guaranteed that a matching set of nuts and bolts will hold together a table just as well as they will hold together a chair, or a piece of metal machinery, or the engine of your car.When we place together a promoter with a new gene of interest we do not yet have these compatibility rules to know how a promoter will work with this gene of interest. We, as synthetic biologists, still don’t know that one promoter/gene will work the same way in one strain of E. coli as they will in another E. coli. For synthetic biology to truly emerge as a field that can capitalize in industry, we must find novel ways of engineering biological systems reliably using our own standard parts.
