单细胞测量质粒拷贝数和启动子活性。

Synthetic Biology Center, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.

Biosystems and Biomaterials Division, Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD, USA.

Nat Commun. 2021 Mar 5;12(1):1475. doi: 10.1038/s41467-021-21734-y.

Accurate measurements of promoter activities are crucial for predictably building genetic systems. Here we report a method to simultaneously count plasmid DNA, RNA transcripts, and protein expression in single living bacteria. From these data, the activity of a promoter in units of RNAP/s can be inferred. This work facilitates the reporting of promoters in absolute units, the variability in their activity across a population, and their quantitative toll on cellular resources, all of which provide critical insights for cellular engineering.

准确测量启动子活性对于可预测地构建遗传系统至关重要。在这里，我们报告了一种在单个活细菌中同时计数质粒 DNA、RNA 转录物和蛋白质表达的方法。根据这些数据，可以推断启动子的活性以 RNA 聚合酶/秒为单位。这项工作有助于以绝对单位报告启动子、它们在群体中的活性变化，以及它们对细胞资源的定量影响，所有这些都为细胞工程提供了关键的见解。