ArticleNature protocols2021
SorTn-seq: a high-throughput functional genomics approach to discovering regulators of bacterial gene expression.
Article in Nature protocols, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- High-throughput transposon sequencing identifies HprR as a key regulator of the LEE operon and virulence inVirulence · 2026Article
- Defining the essential genome of diverse phages with phage Tn-seq.Nature microbiology · 2026Article
- Applications of transposon-insertion sequencing for understanding bacterial physiology.Microbial genomics · 2026Review
- How Klebsiella pneumoniae controls its virulence.PLoS pathogens · 2025Review
- Genome-wide identification of bacterial genes contributing to nucleus-forming jumbo phage infection.Nucleic acids research · 2025Article
- Review
- CRISPR-Cas immunity is repressed by the LysR-type transcriptional regulator PigU.Nucleic acids research · 2024Article
- Identification ofJournal of bacteriology · 2023Article
- High-throughput functional genomics: A (myco)bacterial perspective.Molecular microbiology · 2023Review
- Amoxicillin-resistanteLife · 2022Article
- Antimicrobial resistance: progress and challenges in antibiotic discovery and anti-infective therapy.Microbial biotechnology · 2022Article
- The Rsm (Csr) post-transcriptional regulatory pathway coordinately controls multiple CRISPR-Cas immune systems.Nucleic acids research · 2021Article
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Authors and funding
4 authors.
Funding
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Abstract
We recently developed a high-throughput functional genomics approach, named 'SorTn-seq', to identify factors affecting expression of any gene of interest in bacteria. Our approach facilitates high-throughput screening of complex mutant pools, a task previously hindered by a lack of suitable techniques. SorTn-seq combines high-density, Tn5-like transposon mutagenesis with fluorescence-activated cell sorting of a strain harboring a promoter-fluorescent reporter fusion, to isolate mutants with altered gene expression. The transposon mutant pool is sorted into different bins on the basis of fluorescence, and mutants are deep-sequenced to identify transposon insertions. DNA is prepared for sequencing by using commercial kits augmented with custom primers, enhancing ease of use and reproducibility. Putative regulators are identified by comparing the number of insertions per genomic feature in the different sort bins, by using existing bioinformatic pipelines and software packages. SorTn-seq can be completed in 1-2 weeks and requires general microbiology skills and basic flow cytometry experience.
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