A liquid handling platform for standardised quantification of cell-free enzymatic activity encoded by antimicrobial resistance genes
File(s)
Published version (advance access)
Author(s)
Bergum, Molly
Suthakaran, Sahana
Martin, Bethany
Sutton, J Mark
Moore, Simon
Type
Journal Article
Abstract
Antimicrobial resistance (AMR) is a growing global threat to human health, and rapid methods for characterising emerging antimicrobial resistance genes (ARGs) are needed. Here, we develop a semi-automated workflow using cell-free gene expression (CFE) systems to measure the activity of two ARGs encoded on plasmid DNA that produce rifampicin-inactivating and gentamicin-inactivating enzymes. We validated the use of a small benchtop Myra liquid handling system compared to manual pipetting, with no statistical differences observed. After optimising the pre-incubation time of ARGs and dispensing protocol, expression of aac(3)-IIa increased the half-maximal inhibition concentration (IC50) of gentamicin by over 150-fold, while arr-3 increased the IC50 of rifampicin by approximately 20-fold compared to controls. This methodology for rapid, semi-automated ARG characterisation offers a strategy to combat AMR by assessing novel ARGs identified through genomic surveillance or profiling activity of new or derivative antibiotics.
Date Issued
2026-08-27
Date Acceptance
2026-08-19
Citation
Synthetic Biology, 2026, 11 (1)
ISSN
1939-7267
Publisher
Oxford University Press
Journal / Book Title
Synthetic Biology
Volume
11
Issue
1
Copyright Statement
© The Author(s) 2026. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
License URL
Identifier
10.1093/synbio/ysag010
Subjects
cell-free gene expression
antimicrobial resistance
liquid handling
automation
aminoglycoside resistance
rifampicin resistance
Publication Status
Published online
Article Number
ARTN ysag010
Date Publish Online
2026-08-27