biostudies-literatureUnknownBasta DWNIDDK NIH HHSNIAID NIH HHSTransposon insertion sequencing (Tn-seq) is a powerful method for genome-scale functional genetics in bacteria. However, its effectiveness is often limited by a lack of mutant diversity, caused by either inefficient transposon delivery or stochastic loss of mutants due to population bottlenecks. Here, we introduce "InducTn-seq", which leverages inducible mutagenesis for temporal control of transposition. InducTn-seq generates millions of transposon mutants from a single colony, enabling the sensitive detection of subtle fitness defects and transforming binary classifications of gene essentiality into a quantitative fitness measurement across both essential and non-essential genes. Using a mouse model of infectious colitis, we show that InducTn-seq bypasses a highly restrictive host bottlenbioRxiv : the preprint server for biology2024.05.21.595064https://www.ebi.ac.uk/biostudies/studies/S-EPMC11142078biostudies-literatureInducible transposon mutagenesis for genome-scale forward genetics.PMC11142078F31 AI156949R01 AI042347T32 DK007477P30 DK034854Waldor MKBasta DWHotinger JAHullahalli KSullivan EJCampbell IWfalseInducible transposon mutagenesis for genome-scale forward genetics.Transposon insertion sequencing (Tn-seq) is a powerful method for genome-scale functional genetics in bacteria. However, its effectiveness is often limited by a lack of mutant diversity, caused by either inefficient transposon delivery or stochastic loss of mutants due to population bottlenecks. Here, we introduce "InducTn-seq", which leverages inducible mutagenesis for temporal control of transposition. InducTn-seq generates millions of transposon mutants from a single colony, enabling the sensitive detection of subtle fitness defects and transforming binary classifications of gene essentiality into a quantitative fitness measurement across both essential and non-essential genes. Using a mouse model of infectious colitis, we show that InducTn-seq bypasses a highly restrictive host bottlen2024-01-01T00:00:00Z2024 May2026-04-15T03:32:54.905Z2026-04-15T03:15:39.84ZS-EPMC111420783882632510.1101/2024.05.21.595064