Edwin: a robotic platform for automated RNA extraction and analysis during reporter gene-based dynamic characterisation of bacterial promoters

Aitor De Las Heras, Weike Xiao, Vlastimil Srsen, Alistair Elfick

Research output: Contribution to journalArticlepeer-review

Abstract

Characterization of gene expression is a central tenet of the synthetic
biology design cycle. Sometimes it requires high-throughput approaches
which allow quantification of the gene expression of different elements in
diverse conditions. Recently, several large–scale studies have highlighted
the importance of post-transcriptional regulation mechanisms and their
impact on correlations between mRNA and protein abundance. Here, we
introduce Edwin, a robotic workstation, which 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 E.coli strains carrying different promoter/gfp fusions. We
validated Edwin’s abilities: i) preparation of custom cultures of microbial
cells and ii) 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 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.
Original languageEnglish
Pages (from-to)50-62
JournalJournal of Laboratory Automation
Volume22
Issue number1
Early online date17 Jun 2016
DOIs
Publication statusE-pub ahead of print - 17 Jun 2016

Keywords

  • Synthetic Biology
  • High-throughput robotics
  • gene regulation
  • laboratory automation

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