Inducible transposon mutagenesis for genome-scale forward genetics

Data from transposon insertion sequencing using an inducible mutagenesis system, along with related microbial genome sequencing. 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 bottleneck to generate a diverse transposon mutant population from the cells that initiate infection, revealing the role of oxygen-related metabolic plasticity in pathogenesis. Overall, InducTn-seq overcomes the limitations of traditional Tn-seq, unlocking new possibilities for genome-scale forward genetic screens in bacteria.

本数据集涵盖采用诱导型诱变系统的转座子插入测序（transposon insertion sequencing, Tn-seq）相关数据，以及配套的微生物基因组测序数据。转座子插入测序（Tn-seq）是一种可用于细菌全基因组规模功能遗传学研究的强有力手段。然而，其应用效果常受限于突变体多样性不足，该问题可归因于转座子递送效率低下，或是种群瓶颈引发的突变体随机丢失。本研究提出了诱导型Tn-seq（InducTn-seq），该方法借助诱导型诱变系统实现转座过程的时间精准调控。诱导型Tn-seq可从单个细菌菌落中构建出数百万个转座子突变体，能够灵敏检测出细微的生长适合度缺陷，并将基因必需性的二元分类模式转化为针对必需与非必需基因的定量生长适合度评估体系。借助感染性结肠炎小鼠模型，本研究证实诱导型Tn-seq可突破严苛的宿主种群瓶颈，从启动感染的细菌细胞中构建出多样性丰富的转座子突变体群体，进而揭示了氧相关代谢可塑性在病原菌致病过程中的关键作用。综上，诱导型Tn-seq克服了传统Tn-seq的诸多局限，为细菌全基因组规模的正向遗传筛选开辟了全新的研究可能。