Single-cell measurement of plasmid copy number and promoter activity
Bin Shao, Jayan Rammohan, Daniel A. Anderson, Nina Alperovich, David Ross, Christopher A. Voigt
Abstract
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.
Topics & Concepts
PlasmidPromoter activityPromoterBiologyComputational biologyDNAGeneticsRNAPopulationGeneGene expressionDemographySociologyBacterial Genetics and BiotechnologyGene Regulatory Network AnalysisCRISPR and Genetic Engineering