Edwin.
Characterization of gene expression is a central tenet of the synthetic biology design cycle. Sometimes it requires high-throughput approaches that allow quantification of the gene expression of different elements in diverse conditions. Recently, several large-scale studies have highlighted the importance of posttranscriptional regulation mechanisms and their impact on correlations between mRNA and protein abundance. Here, we introduce Edwin, a robotic workstation that enables the automated propagation of microbial cells and the dynamic characterization of gene expression. We developed an automated procedure that integrates customized RNA extraction and analysis into the typical high-throughput characterization of reporter gene expression. To test the system, we engineered Escherichia coli strains carrying different promoter/ gfp fusions. We validated Edwin's abilities: (1) preparation of custom cultures of microbial cells and (2) dynamic quantification of fluorescence signal and bacterial growth and simultaneous RNA extraction and analysis at different time points. We confirmed that RNA obtained during this automated process was suitable for use in qPCR analysis. Our results established that Edwin is a powerful platform for the automated analysis of microbial gene expression at the protein and RNA level. This platform could be used in a high-throughput manner to characterize not only natural regulatory elements but also synthetic ones.
1 Institute for Bioengineering, School of Engineering, University of Edinburgh, Edinburgh, UK.|2 SynthSys Research Centre, University of Edinburgh, Edinburgh, UK.
Aitor
A
de Las Heras
1 Institute for Bioengineering, School of Engineering, University of Edinburgh, Edinburgh, UK.
Weike
W
Xiao
1 Institute for Bioengineering, School of Engineering, University of Edinburgh, Edinburgh, UK.
Vlastimil
V
Sren
1 Institute for Bioengineering, School of Engineering, University of Edinburgh, Edinburgh, UK.|2 SynthSys Research Centre, University of Edinburgh, Edinburgh, UK.
Alistair
A
Elfick
D057205:Automation, Laboratory / Q000295:instrumentation / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics*; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000032:analysis* / Q000302:isolation & purification*; D012371:Robotics / Q000295:instrumentation / Q000379:methods
D012329:RNA, Bacterial; D049452:Green Fluorescent Proteins
gene regulation; high-throughput robotics; laboratory automation; synthetic biology