2026· Methods in molecular biology· Vol 3055, pp.
53-65
· 0 citations
Medicine
TL;DR
This chapter provides a detailed, step-by-step protocol for implementing a conditional plasmid system that enables efficient, markerless gene deletion in FNA strains and provides a powerful and adaptable tool for advancing genetic studies in this genetically recalcitrant subspecies.
This chapter presents a step-by-step protocol for designing sgRNAs, constructing CRISPRi plasmids, transforming F. nucleatum ATCC 23726, and evaluating gene silencing phenotypes, using the nonessential gene ftsW, which encodes a protein required for peptidoglycan synthesis and cell division, as a model target.
Shiqi Xu, B. C., Kexin Tan et al.· Methods in molecular biology· 0 citations
A counterselection system based on pheS∗ that enables markerless gene deletion in P. gingivalis is established and provides a practical and broadly applicable framework for advanced genetic manipulation in this important oral pathobiont.
Hey-Min Kim, M. E. Davey, Manda Yu· Microbiology spectrum· 0 citations
An inducible genome-editing framework exploiting the endogenous Cas9 system of Apilactobacillus kunkeei, a key member of the honeybee microbiota, is established and the toolkit available for harnessing endogenous CRISPR-Cas systems in genetically recalcitrant, non-model gram-positive bacteria is expanded.
Mahesh S. Iyer, Erik Hagström, Kristina Näslund et al.· Applied and Environmental Mi...· 0 citations
A second-generation genetic toolbox based on leucine auxotrophy that enables antibiotic-free positive selection in Mycobacterium abscessus and significantly reduces dependency on antibiotics for genetic manipulation, aligning with the goals of sustainable research and offering new opportunities for studying this clinic...
Tizian Griesser, P. Selchow, Peter Sander· Applied and Environmental Mi...· 0 citations
This chapter describes strategies to evade RM defenses and improve transformation efficiencies across diverse Fusobacterium lineages, employing a sequence-based "RM-silencing" method that has enabled successful delivery of replicative plasmids, linear recombination templates, and transposon cassettes into previously in...
Martha A. Zepeda-Rivera, Elsa F. McMahon, Kaitlyn N. Lewis et al.· Methods in molecular biology· 0 citations
Improved the previously developed tool by designing a single-plasmid variant instead of the two-plasmid one and by adapting the tetracycline resistance gene as an additional selective marker in L. lactis, significantly simplifies and accelerates the delivery of the required genetic material into the chromosome.
P. Pechenov, O. N. Levin, D. V. Verevochnikov et al.· Microbiology· 0 citations
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