BIOFAB International Open Facility Advancing Biotechnology, Emeryville, California, USA.
Nat Methods. 2013 Apr;10(4):354-60. doi: 10.1038/nmeth.2404. Epub 2013 Mar 10.
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 ∼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.
无法可靠地预测新的遗传元件组合的定量行为限制了生物系统的合理工程设计。我们开发了一种用于控制大肠杆菌转录和翻译起始的基因元件表达盒架构：转录元件编码一个共同的 mRNA 起始点，翻译元件使用在许多天然系统中发现的重叠遗传模体。我们按照这些定义设计了组成型和阻遏物调节启动子文库以及翻译起始元件。我们测量了每个文库的活性分布，并选择了共同导致在 1000 倍观察到的动态范围内表达的元件。我们研究了所有经过策展的元件的组合，证明了任意基因的表达都可以可靠地控制在目标表达窗口的两倍以内，可靠性约为 93%。我们期望这里验证的遗传元件定义可以被集体扩展，以创建支持在基因组范围内可靠地进行基因表达正向工程的公共领域标准生物部件集合。
