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An automated platform for accelerating and focusing adaptive laboratory evolution

Ruppen, Peter ; Bahls, Maximilian Ole ; Gerlt, Michael Sebastian LU orcid ; Edelmann, Martin Peter ; Roberts, Tania Michelle ; Marlière, Philippe and Panke, Sven (2026) In Metabolic Engineering 94. p.241-251
Abstract

The rate of change in adaptive laboratory evolution (ALE), in which a population of microorganisms is continuously cultivated under a specific selective pressure, is controlled by the cellular mutagenesis rate and the randomness of where in the genetic material mutations are introduced. The constant selection pressure makes it a crucial, yet slow, method in developing microorganisms with novel phenotypes for which a rational engineering pathway is either too complex or unknown. A variety of targeted genome editing methods to accelerate evolution and facilitate the engineering of complex novel traits are available. However, these protocols require (nearly) as many successive transformation steps as loci they target, leaving the actual... (More)

The rate of change in adaptive laboratory evolution (ALE), in which a population of microorganisms is continuously cultivated under a specific selective pressure, is controlled by the cellular mutagenesis rate and the randomness of where in the genetic material mutations are introduced. The constant selection pressure makes it a crucial, yet slow, method in developing microorganisms with novel phenotypes for which a rational engineering pathway is either too complex or unknown. A variety of targeted genome editing methods to accelerate evolution and facilitate the engineering of complex novel traits are available. However, these protocols require (nearly) as many successive transformation steps as loci they target, leaving the actual engineering process quite labor-intense, cumbersome, and at odds with the continuous nature of ALE. Here, we provide a fully integrated microfluidic platform that automates and accelerates bacterial transformation by electroporation to the mere push of a button. We demonstrate the functionality and effect by using oligonucleotide-directed mutagenesis in an ALE experiment to accelerate the engineering of riboflavin prototrophy into Escherichia coli .

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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Automation, E. coli, Electroporation, Genome editing, Medium exchange, Microfluidics, Transformation
in
Metabolic Engineering
volume
94
pages
11 pages
publisher
Academic Press
external identifiers
  • scopus:105026661886
  • pmid:41455546
  • pmid:41455546
ISSN
1096-7176
DOI
10.1016/j.ymben.2025.12.007
language
English
LU publication?
yes
additional info
Publisher Copyright: © 2025 The Authors.
id
406c88d4-6e26-4722-a7e6-2e64665a8e3c
date added to LUP
2026-01-05 21:54:49
date last changed
2026-01-19 14:48:49
@article{406c88d4-6e26-4722-a7e6-2e64665a8e3c,
  abstract     = {{<p>The rate of change in adaptive laboratory evolution (ALE), in which a population of microorganisms is continuously cultivated under a specific selective pressure, is controlled by the cellular mutagenesis rate and the randomness of where in the genetic material mutations are introduced. The constant selection pressure makes it a crucial, yet slow, method in developing microorganisms with novel phenotypes for which a rational engineering pathway is either too complex or unknown. A variety of targeted genome editing methods to accelerate evolution and facilitate the engineering of complex novel traits are available. However, these protocols require (nearly) as many successive transformation steps as loci they target, leaving the actual engineering process quite labor-intense, cumbersome, and at odds with the continuous nature of ALE. Here, we provide a fully integrated microfluidic platform that automates and accelerates bacterial transformation by electroporation to the mere push of a button. We demonstrate the functionality and effect by using oligonucleotide-directed mutagenesis in an ALE experiment to accelerate the engineering of riboflavin prototrophy into Escherichia coli .</p>}},
  author       = {{Ruppen, Peter and Bahls, Maximilian Ole and Gerlt, Michael Sebastian and Edelmann, Martin Peter and Roberts, Tania Michelle and Marlière, Philippe and Panke, Sven}},
  issn         = {{1096-7176}},
  keywords     = {{Automation; E. coli; Electroporation; Genome editing; Medium exchange; Microfluidics; Transformation}},
  language     = {{eng}},
  pages        = {{241--251}},
  publisher    = {{Academic Press}},
  series       = {{Metabolic Engineering}},
  title        = {{An automated platform for accelerating and focusing adaptive laboratory evolution}},
  url          = {{http://dx.doi.org/10.1016/j.ymben.2025.12.007}},
  doi          = {{10.1016/j.ymben.2025.12.007}},
  volume       = {{94}},
  year         = {{2026}},
}