ARTIST: High-Resolution Genome-Wide Assessment of Fitness Using Transposon-Insertion Sequencing

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Transposon insertion sequencing (TIS) is a powerful method that couples high-density transposon mutagenesis with next-generation sequencing to comprehensively assess the fitness of thousands of transposon mutants across a genome. TIS is an extremely flexible technique that has been used to define genomic loci required for bacterial growth and survival in a variety of species and in many different growth conditions, including during host infection. However, there remain several important limitations to current TIS analysis methods. First, TIS data are not routinely normalized for the impact of experimental variability; second, most analyses are restricted to annotated loci and do not completely exploit the richness of TIS datasets; finally, TIS analysis methods are not easily accessible to most biologists. Here we present a pipeline—ARTIST—that addresses these issues and will transform TIS-based studies. We used ARTIST to conduct robust analyses of Mycobacterium tuberculosis and Vibrio cholerae in vivo TIS datasets and comprehensively defined the genetic requirements of these pathogens for host infection. The ARTIST pipeline will make TIS analysis accessible to many researchers and greatly enhance the rigor of and insights gained from TIS studies in a wide range of microorganisms.

Vyšlo v časopise: ARTIST: High-Resolution Genome-Wide Assessment of Fitness Using Transposon-Insertion Sequencing. PLoS Genet 10(11): e32767. doi:10.1371/journal.pgen.1004782
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1004782

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