Screening conditions and constructs for attempted genetic transformation of C. elegans by Agrobacterium

Manipulating gene expression within a model organism is important for reverse genetic experimentation, and while techniques to generate transgenic C. elegans are available, they are optimised for creating individual lines. The ability to create libraries of genetically modified animals using C. elegans as a model would make new types of experiments possible and would speed up studies of animal physiology. Here, we describe a range of constructs designed to establish a high-throughput method of C. elegans transformation mediated by gene transfer from Agrobacterium. We demonstrate that C. elegans are able to survive on Agrobacterium as a sole food source, and screen conditions for Agrobacterium-mediated transformation in this organism. While we do not achieve routine gene transfer from Agrobacterium to C. elegans, we suggest that this technique has potential following further optimization. The success of the approach would enable rapid and high-throughput transformation of C. elegans, providing an improvement on currently available methods. Here we provide details of optimization conditions tested, and a useful resource of T-binary constructs for use by the scientific community.

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Journal
PLoS ONE
Published
2025-05-27
DOI
https://doi.org/10.1371/journal.pone.0325060
Primary Topic
Genetics, Aging, and Longevity in Model Organisms
Type
article
Field-Weighted Citation Impact
0.00

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article

Screening conditions and constructs for attempted genetic transformation of C. elegans by Agrobacterium

Karen S. Sarkisyan, André EX Brown, Eleanor C. Warren
PLoS ONE
Genetics, Aging, and Longevity in Model Organisms
article

Screening conditions and constructs for attempted genetic transformation of C. elegans by Agrobacterium

Karen S. Sarkisyan, André EX Brown, Eleanor C. Warren
article en

Abstract

Manipulating gene expression within a model organism is important for reverse genetic experimentation, and while techniques to generate transgenic C. elegans are available, they are optimised for creating individual lines. The ability to create libraries of genetically modified animals using C. elegans as a model would make new types of experiments possible and would speed up studies of animal physiology. Here, we describe a range of constructs designed to establish a high-throughput method of C. elegans transformation mediated by gene transfer from Agrobacterium. We demonstrate that C. elegans are able to survive on Agrobacterium as a sole food source, and screen conditions for Agrobacterium-mediated transformation in this organism. While we do not achieve routine gene transfer from Agrobacterium to C. elegans, we suggest that this technique has potential following further optimization. The success of the approach would enable rapid and high-throughput transformation of C. elegans, providing an improvement on currently available methods. Here we provide details of optimization conditions tested, and a useful resource of T-binary constructs for use by the scientific community.

PLoS ONEVol. 20(5)
MRC London Institute of Medical Sciences (GB), Imperial College London (GB)
Medical Research Council, Medical Research Foundation
Openalex Percentile: Top 90%
Genetics, Aging, and Longevity in Model Organisms
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