The practice of engineering biology now depends on the ad hoc reuse of genetic elements whose precise activities vary across changing contexts. Methods are lacking for researchers to affordably coordinate the quantification and analysis of part performance across varied environments, as needed to identify, evaluate and improve problematic part types. We developed an easy-to-use analysis of variance (ANOVA) framework for quantifying the performance of genetic elements. For proof of concept, we assembled and analyzed combinations of prokaryotic transcription and translation initiation elements in Escherichia coli. We determined how estimation of part activity relates to the number of unique element combinations tested, and we show how to estimate expected ensemble-wide part activity from just one or two measurements. We propose a new statistic, biomolecular part 'quality', for tracking quantitative variation in part performance across changing contexts.
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We thank N. Hillson, V. Rhodius and C. Smolke for comments and discussion, and F. St-Pierre (Stanford University) for the plasmid pIT-KL-I52002. We acknowledge support from the National Science Foundation to the BIOFAB (award EEC 0946510). G.C. acknowledges support from the Human Frontier Science Program (LT000873/2011-l) and the Bettencourt Schueller foundation. A.P.A. and D.E. acknowledge support from the Synthetic Biology Engineering Research Center under National Science Foundation grant 04-570/0540879. J.C.G. acknowledges financial support from the Portuguese Fundação para a Ciência e a Tecnologia (SFRH/BD/47819/2008). This work was conducted at the Joint BioEnergy Institute supported by the Office of Science, Office of Biological and Environmental Research of the US Department of Energy, under contract DE-AC02-05CH11231.
The authors declare no competing financial interests.
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Mutalik, V., Guimaraes, J., Cambray, G. et al. Quantitative estimation of activity and quality for collections of functional genetic elements. Nat Methods 10, 347–353 (2013). https://doi.org/10.1038/nmeth.2403
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