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High-throughput chemical genomic screening: a step-by-step workflow from plate to phenotype

Title: High-throughput chemical genomic screening: a step-by-step workflow from plate to phenotype
Authors: Georgia Williams; Huda Ahmad; Susan Sutherland; James Haycocks; Sam Benedict; Adam J. Hart; Hannah H. Doherty; Rudi Sullivan; Micheal Alao; Xuyu Ma; Qianhui Xu; Jack Bryant; Monika Glinkowska; Peter Banks; Patrick Moynihan; Mathew T. Milner; Danesh Moradigaravand; Manuel Banzhaf
Source: mSystems, Vol 10, Iss 12 (2025)
Publisher Information: American Society for Microbiology, 2025.
Publication Year: 2025
Collection: LCC:Microbiology
Subject Terms: chemical genomics; bioinformatics; genomics; systems microbiology; Microbiology; QR1-502
Description: ABSTRACT High-throughput chemical genomics uses phenotypic profiling of strain libraries under defined chemical and environmental conditions to identify gene functions. This approach enables the mapping of biological pathways and can potentially highlight drug targets. Chemical genomic data sets have been springboards for numerous hypothesis-driven research projects, with direct implications for antimicrobial resistance and clinical outcomes. High-throughput phenotypic profiles are valuable tools for enriching microbial sequence data with functional annotations and benefiting the broader scientific community. This work provides a step-by-step guide for conducting chemical genomics screens from start to finish.IMPORTANCEChemical genomic screening is a powerful systems biology approach for linking gene function to phenotype under diverse chemical and environmental stressors. However, its broader use in microbial research has been limited by the lack of standardized, reproducible workflows. Our study introduces a scalable, end-to-end protocol that integrates experimental, imaging, and computational steps into a cohesive framework for high-throughput screening across a range of microbial species. This enables researchers to generate consistent, high-quality phenotypic data suitable for large-scale analyses. The protocol supports systematic exploration of gene-environment interactions, microbial stress responses, and antimicrobial resistance. Its adaptability and troubleshooting guidance make it especially useful for groups working in microbiome research, synthetic biology, and microbial community studies. By bridging benchwork and computational analysis, this workflow expands the technical toolkit available to microbial systems biologists. Our work contributes to the development of robust methods for functional genomics and supports the core mission of mSystems to advance microbial systems biology.
Document Type: article
File Description: electronic resource
Language: English
ISSN: 2379-5077
Relation: https://doaj.org/toc/2379-5077
DOI: 10.1128/msystems.00885-25
Access URL: https://doaj.org/article/8942e4bfd6fc49a78f7ffb29a9e91b8d
Accession Number: edsdoj.8942e4bfd6fc49a78f7ffb29a9e91b8d
Database: Directory of Open Access Journals