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Review

Essential Genes Discovery in Microorganisms by Transposon-Directed Sequencing (Tn-Seq): Experimental Approaches, Major Goals, and Future Perspectives

by
Gemma Fernández-García
,
Paula Valdés-Chiara
,
Patricia Villazán-Gamonal
,
Sergio Alonso-Fernández
and
Angel Manteca
*
Department of Functional Biology, Microbiology Area, IUOPA and ISPA, Faculty of Medicine, Universidad de Oviedo, 33006 Oviedo, Spain
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2024, 25(20), 11298; https://doi.org/10.3390/ijms252011298
Submission received: 23 September 2024 / Revised: 18 October 2024 / Accepted: 19 October 2024 / Published: 21 October 2024
(This article belongs to the Special Issue Current Advances and Perspectives in Microbial Genetics and Genomics)
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Abstract

Essential genes are crucial for microbial viability, playing key roles in both the primary and secondary metabolism. Since mutations in these genes can threaten organism viability, identifying them is challenging. Conditionally essential genes are required only under specific conditions and are important for functions such as virulence, immunity, stress survival, and antibiotic resistance. Transposon-directed sequencing (Tn-Seq) has emerged as a powerful method for identifying both essential and conditionally essential genes. In this review, we explored Tn-Seq workflows, focusing on eubacterial species and some yeast species. A comparison of 14 eubacteria species revealed 133 conserved essential genes, including those involved in cell division (e.g., ftsA, ftsZ), DNA replication (e.g., dnaA, dnaE), ribosomal function, cell wall synthesis (e.g., murB, murC), and amino acid synthesis (e.g., alaS, argS). Many other essential genes lack clear orthologues across different microorganisms, making them specific to each organism studied. Conditionally essential genes were identified in 18 bacterial species grown under various conditions, but their conservation was low, reflecting dependence on specific environments and microorganisms. Advances in Tn-Seq are expected to reveal more essential genes in the near future, deepening our understanding of microbial biology and enhancing our ability to manipulate microbial growth, as well as both the primary and secondary metabolism.
Keywords: Tn-Seq; transposon; essential genes Tn-Seq; transposon; essential genes

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MDPI and ACS Style

Fernández-García, G.; Valdés-Chiara, P.; Villazán-Gamonal, P.; Alonso-Fernández, S.; Manteca, A. Essential Genes Discovery in Microorganisms by Transposon-Directed Sequencing (Tn-Seq): Experimental Approaches, Major Goals, and Future Perspectives. Int. J. Mol. Sci. 2024, 25, 11298. https://doi.org/10.3390/ijms252011298

AMA Style

Fernández-García G, Valdés-Chiara P, Villazán-Gamonal P, Alonso-Fernández S, Manteca A. Essential Genes Discovery in Microorganisms by Transposon-Directed Sequencing (Tn-Seq): Experimental Approaches, Major Goals, and Future Perspectives. International Journal of Molecular Sciences. 2024; 25(20):11298. https://doi.org/10.3390/ijms252011298

Chicago/Turabian Style

Fernández-García, Gemma, Paula Valdés-Chiara, Patricia Villazán-Gamonal, Sergio Alonso-Fernández, and Angel Manteca. 2024. "Essential Genes Discovery in Microorganisms by Transposon-Directed Sequencing (Tn-Seq): Experimental Approaches, Major Goals, and Future Perspectives" International Journal of Molecular Sciences 25, no. 20: 11298. https://doi.org/10.3390/ijms252011298

APA Style

Fernández-García, G., Valdés-Chiara, P., Villazán-Gamonal, P., Alonso-Fernández, S., & Manteca, A. (2024). Essential Genes Discovery in Microorganisms by Transposon-Directed Sequencing (Tn-Seq): Experimental Approaches, Major Goals, and Future Perspectives. International Journal of Molecular Sciences, 25(20), 11298. https://doi.org/10.3390/ijms252011298

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