Journal
NATURE METHODS
Volume 10, Issue 4, Pages 354-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.2404
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Funding
- US National Science Foundation [EEC 0946510]
- Portuguese Fundacao para a Ciencia e a Tecnologia (FCT) [SFRH/BD/47819/2008]
- Human Frontier Science Program [LT000873/2011-l]
- Bettencourt Schueller Foundation
- Synthetic Biology Engineering Research Center under National Science Foundation [04-570/0540879]
- Office of Science, Office of Biological and Environmental Research, US Department of Energy [DE-AC02-05CH11231]
- US National Science Foundation [EEC 0946510]
- Portuguese Fundacao para a Ciencia e a Tecnologia (FCT) [SFRH/BD/47819/2008]
- Human Frontier Science Program [LT000873/2011-l]
- Bettencourt Schueller Foundation
- Synthetic Biology Engineering Research Center under National Science Foundation [04-570/0540879]
- Office of Science, Office of Biological and Environmental Research, US Department of Energy [DE-AC02-05CH11231]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/47819/2008] Funding Source: FCT
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An inability to reliably predict quantitative behaviors for novel combinations of genetic elements limits the rational engineering of biological systems. We developed an expression cassette architecture for genetic elements controlling transcription and translation initiation in Escherichia coli: transcription elements encode a common mRNA start, and translation elements use an overlapping genetic motif found in many natural systems. We engineered libraries of constitutive and repressor-regulated promoters along with translation initiation elements following these definitions. We measured activity distributions for each library and selected elements that collectively resulted in expression across a 1,000-fold observed dynamic range. We studied all combinations of curated elements, demonstrating that arbitrary genes are reliably expressed to within twofold relative target expression windows with similar to 93% reliability. We expect the genetic element definitions validated here can be collectively expanded to create collections of public-domain standard biological parts that support reliable forward engineering of gene expression at genome scales.
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