Tn-seq; high-throughput parallel sequencing for fitness and genetic interaction studies in microorganisms
By T. van Opijnen, K. Bodi, A. Camilli. Nature Methods, 2009
918 citations recorded by the source corpus.
Biological pathways are structured in complex networks of interacting genes. Solving the architecture of such networks may provide valuable information, such as how microorganisms cause disease. Here we present a method (Tn-seq) for accurately determining quantitative genetic interactions on a genome-wide scale in microorganisms. Tn-seq is based on the assembly of a saturated Mariner transposon insertion library. After library selection, changes in frequency of each insertion mutant are determined by sequencing the flanking regions en masse. These changes are used to calculate each mutant's fitness. Using this approach, we determined fitness for each gene of Streptococcus pneumoniae, a causative agent of pneumonia and meningitis. A genome-wide screen for genetic interactions of five query genes identified both alleviating and aggravating interactions that could be divided into seven distinct categories. Owing to the wide activity of the Mariner transposon, Tn-seq has the potential to contribute to the exploration of complex pathways across many different species.
Subfields: Ecology, Molecular Biology
View Paper: https://www.semanticscholar.org/paper/c506062407cebe8cb4e7cd709e3160cc3d3ca2ed
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Citations in the corpus
- Intracellular staphylococcus aureus: Live-in and let die (2012, positive): “However, novel high-throughput sequencing technologies of transposon insertions (Gawronski et al., 2009; van Opijnen et al., 2009) open up new possibilities for analysis of genome-wide mutant libraries of clinically relevant strains.”
- DEG 10, an update of the database of essential genes that includes both protein-coding genes and noncoding genomic elements (2013, background): “Gallagher,L.A., Shendure,J. and Manoil,C. (2011) Genome-scale identification of resistance functions in Pseudomonas aeruginosa using Tn-seq.”