Dual transposon sequencing profiles the genetic interaction landscape in bacteria

• 成果类型: Article
• 署名作者: Zik, Justin J.; Price, Morgan N.; Mayra, Keisha Hanifa Alma; Santoso, Audrey A.; Arkin, Adam P.; Deutschbauer, Adam M.; Sham, Lok-To
• 署名单位: National University of Singapore; National University of Singapore; United States Department of Energy (DOE); Lawrence Berkeley National Laboratory; University of California System; University of California Berkeley; University of California System; University of California Berkeley
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-10095
• DOI: 10.1126/science.adt7685
• 发表日期: 2025-09-25
• 关键词: streptococcus-pneumoniae; escherichia-coli; high-throughput; cell-shape; wild-type; virulence; metabolism; dna; triphosphatase; biosynthesis

摘要

Gene redundancy complicates systematic characterization of gene function as single-gene deletions may not produce discernible phenotypes. We report dual transposon sequencing (dual Tn-seq), a platform for assaying the fitness of a comprehensive double mutant pool in parallel. Dual Tn-seq couples random barcode transposon site sequencing with the Cre-lox system, enabling deep sampling of 73% of the 1.3 million possible double gene deletions in Streptococcus pneumoniae. The genetic interactions identified span a wide range of biochemical processes, revealing new factors in presumably well-studied pathways, exemplified by a cytidine triphosphate synthase PyrJ. Moreover, this approach should permit further investigation of growth condition-specific genetic interactions. Because dual Tn-seq does not require the construction of a large array of single mutants, it should be readily adaptable to various microorganisms.