Establishment and improvement of genetic manipulation tools for Fusobacterium nucleatum

Abstract

An imbalance in oral microbial homeostasis is significantly associated with the onset and progression of several systemic diseases. Fusobacterium nucleatum, a ubiquitous periodontitis-causing bacterium in the oral cavity, is frequently detected in focal sites and contributes to the pathogenesis of many extraoral diseases, including cancers, cardiovascular diseases, and adverse pregnancy outcomes (APOs). F. nucleatum is one of the few oral anaerobes that can be cultured purely in vitro and is a ‘model species’ for studying the impact of oral health on systemic health. The establishment and development of genetic manipulation tools for F. nucleatum and the construction of pathogenic gene-disrupted strains are important strategies for studying the pathogenicity of F. nucleatum. Here, we review the establishment and development of the genetic manipulation systems for F. nucleatum and summarize the characteristics of various genetic manipulation tools, such as suicide plasmid-based systems for gene inactivation, replicable plasmid-based systems controlling gene expression, and transposon-based random mutagenesis systems. Notably, we summarize and analyze their applications in the study of the pathogenic mechanisms of F. nucleatum. We hope to provide reference information and ideas for future research on genetic manipulation tools and the pathogenic mechanisms of F. nucleatum and other Fusobacterium species.

Keywords

Fusobacterium nucleatum / Genetic manipulation tool / Gene inactivation / Restriction-modification system / Natural competence / Crispr interference

Declaration of Competing Interest

The authors declare no conflict of interest. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

CRediT authorship contribution statement

Zhiwei Guan: Writing - original draft, Conceptualization. Hailong Wang: Writing - review & editing, Funding acquisition, Conceptualization. Qiang Feng: Writing - review & editing, Funding acquisition, Conceptualization.

Acknowledgments

We sincerely thank the scientists, including but not limited to Susan Kinder-Haake, Yiping W Han, Daniel J Slade, and Chenggang Wu, for their contributions to the establishment and improvement of the genetic manipulation system for F. nucleatum. The figures in this review were created in https://BioRender.com. We sincerely thank the foundation support of the National Natural Science Foundation of China (82270980, 82071122), the National Young Scientist Support Foundation (2019), the Major Innovation Projects in Shandong Province (2021SFGC0502), the Oral Microbiome Innovation Team of Shandong Province (2020KJK001), Shandong Province Key Research and Development Program (2021ZDSYS18), and Intramural Joint Program Fund of State Key Laboratory of Microbial Technology (SKLMTIJP-2024-08).

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