Evidence map›Paper›PMID 39843793›Full record

Articlenpj antimicrobials and resistance2023

A cell-free strategy for host-specific profiling of intracellular antibiotic sensitivity and resistance.

Kameshwari Chengan, Charlotte Hind, Maria Stanley, Matthew E Wand, Lakshmeesha K Nagappa, Kevin Howland, Tanith Hanson, Rubén Martín-Escolano, Anastasios D Tsaousis, José A Bengoechea and 3 more

Abstract read
In one paragraph

Article in npj antimicrobials and resistance, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

4 citing papers in PubMed.

  1. Review
  2. Article
  3. Gene circuit-based sensors.Fundamental research · 2025
    Review
  4. Cell-free protein synthesis platforms for accelerating drug discovery.Biotechnology notes (Amsterdam, Netherlands) · 2025
    Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

13 authors.

Kameshwari ChenganSchool of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, CT7 2NJ, United Kingdom.
Charlotte HindTechnology Development Group, Research and Evaluation, UK Health Security Agency, Salisbury, SP4 0JG, United Kingdom.
Maria StanleySchool of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, CT7 2NJ, United Kingdom.
Matthew E WandTechnology Development Group, Research and Evaluation, UK Health Security Agency, Salisbury, SP4 0JG, United Kingdom.
Lakshmeesha K NagappaSchool of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, CT7 2NJ, United Kingdom.
Kevin HowlandSchool of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, CT7 2NJ, United Kingdom.
Tanith HansonSchool of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, CT7 2NJ, United Kingdom.
Rubén Martín-EscolanoSchool of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, CT7 2NJ, United Kingdom.
Anastasios D TsaousisSchool of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, CT7 2NJ, United Kingdom.ORCID http://orcid.org/0000-0002-5424-1905
José A BengoecheaWellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, Belfast, BT9 7BL, United Kingdom.
J Mark SuttonTechnology Development Group, Research and Evaluation, UK Health Security Agency, Salisbury, SP4 0JG, United Kingdom.
Christopher M SmalesSchool of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, CT7 2NJ, United Kingdom.ORCID http://orcid.org/0000-0002-2762-4724
Simon J MooreSchool of Biological and Behavioural Sciences, Queen Mary University of London, London, E1 4NS, United Kingdom. simon.moore@qmul.ac.uk.

Funding

Biochemical Society ERF-0016Royal Society RGS\R1\231113
6 · The paper itself

Abstract

Antimicrobial resistance (AMR) is a pandemic spread across multiple infectious disease-causing microbes. To provide a host-specific tool to study antibiotic susceptibility and resistance, here we develop Klebsiella pneumoniae cell-free gene expression (CFE) systems from laboratory and clinical isolates. Using proteomics, we identify relative differences and unique proteins for these new CFE systems in comparison to an Escherichia coli MG1655 CFE model. Then we profile antimicrobial susceptibility in parallel with whole cells to quantify CFE antibiotic potency. Finally, we apply this native CFE tool to study AMR variants at a proof-of-concept level. Definably we show that RpoB H526L confers a 58-fold increase in CFE resistance to rifampicin-a genotype observed in rifampicin-resistant Mycobacterium tuberculosis clinical isolates. Overall, we provide a cell-free synthetic biology strategy for the profiling of antibiotic sensitivity and resistance from K. pneumoniae. While initial extract processing requires Biosafety Level 2, the CFE system is non-living, suitable for long-term storage and study in a Biosafety Level 1 lab. We anticipate the K. pneumoniae CFE bioassay is advantageous for host-specific antimicrobial testing, the characterisation of intracellular AMR variants and potentially structure-activity relationship studies.

Identifiers

PMID39843793
PMCPMC11721408

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.