ArticleProceedings of the National Academy of Sciences of the United States of America2009
Tracking insertion mutants within libraries by deep sequencing and a genome-wide screen for Haemophilus genes required in the lung.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 246 papers.
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Who cites it
246 citing papers in PubMed, 376 citations in OpenAlex.
- Enhanced in vivo fitness of carbapenem-resistant oprD mutants of Pseudomonas aeruginosa revealed through high-throughput sequencing.Proceedings of the National Academy of Sciences of the United States of America · 2013Trial
- Transposon-based genome editing of industrial microorganisms: advances, challenges, and prospects.Synthetic and systems biotechnology · 2026Review
- Dual roles of genes required for intrinsic resistance to clarithromycin in evasion of killing by serum complement inInfection and immunity · 2026Article
- Genome microbiology at 20 years: who we are and where we are going.DNA research : an international journal for rapid publication of reports on genes and genomes · 2026Article
- Applications of transposon-insertion sequencing for understanding bacterial physiology.Microbial genomics · 2026Review
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- De novo or Salvage? Nucleotide Availability as a Driver of Bacterial Adaptation and Virulence.MicrobiologyOpen · 2026Review
- GeneScanner: profiling genetic variation across bacterial populations.Microbial genomics · 2026Article
- Unravelling the unique essential genes ofMicrobial genomics · 2026Article
- High-throughput characterization of Mycobacterium tuberculosis gene function across diverse conditions.PLoS biology · 2026Article
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- CRISPRi-seq in Haemophilus influenzae reveals genome-wide and medium-specific growth determinants.PLoS pathogens · 2025Article
- Genome-wide identification of essential genes in the invasive Streptococcus anginosus strain.Scientific reports · 2025Article
- Vibrio cholerae motility is associated with inter-animal transmission.Nature communications · 2025Article
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- Landscape of essential growth and fluconazole-resistance genes in the human fungal pathogen Cryptococcus neoformans.PLoS biology · 2025Article
- Inducible transposon mutagenesis identifies bacterial fitness determinants during infection in mice.Nature microbiology · 2025Article
- UnravelingmSystems · 2025Article
- PIMMS-Dash: Accessible analysis, interrogation, and visualisation of high-throughput transposon insertion sequencing (TIS) data.Computational and structural biotechnology journal · 2024Article
186 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Rapid genome-wide identification of genes required for infection would expedite studies of bacterial pathogens. We developed genome-scale "negative selection" technology that combines high-density transposon mutagenesis and massively parallel sequencing of transposon/chromosome junctions in a mutant library to identify mutants lost from the library after exposure to a selective condition of interest. This approach was applied to comprehensively identify Haemophilus influenzae genes required to delay bacterial clearance in a murine pulmonary model. Mutations in 136 genes resulted in defects in vivo, and quantitative estimates of fitness generated by this technique were in agreement with independent validation experiments using individual mutant strains. Genes required in the lung included those with characterized functions in other models of H. influenzae pathogenesis and genes not previously implicated in infection. Genes implicated in vivo have reported or potential roles in survival during nutrient limitation, oxidative stress, and exposure to antimicrobial membrane perturbations, suggesting that these conditions are encountered by H. influenzae during pulmonary infection. The results demonstrate an efficient means to identify genes required for bacterial survival in experimental models of pathogenesis, and this approach should function similarly well in selections conducted in vitro and in vivo with any organism amenable to insertional mutagenesis.
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Registered trials
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.