ArticleJournal of visualized experiments : JoVE2020
Markerless Gene Deletion by Floxed Cassette Allelic Exchange Mutagenesis in Chlamydia trachomatis.
Article in Journal of visualized experiments : JoVE, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
12 citing papers in PubMed.
- Generation of a toxin/antitoxin-based counterselection marker forInfection and immunity · 2025Article
- Development of shuttle vector-based transformation systems for veterinary and zoonotic chlamydiae.Microbiology spectrum · 2025Article
- Article
- COPB1-knockdown induced type I interferon signaling activation inhibitsFrontiers in microbiology · 2025Article
- Article
- Recent advances in genetic systems in obligate intracellular human-pathogenic bacteria.Frontiers in cellular and infection microbiology · 2023Review
- Advances in genetic manipulation ofFrontiers in immunology · 2023Review
- Article
- A Minimal Replicon Enables Efficacious, Species-Specific Gene Deletion in Chlamydia and Extension of Gene Knockout Studies to the Animal Model of Infection Using Chlamydia muridarum.Infection and immunity · 2022Article
- Article
- The inclusion membrane protein IncS is critical for initiation of the Chlamydia intracellular developmental cycle.PLoS pathogens · 2022Article
- Expression and structure of the Chlamydia trachomatis DksA ortholog.Pathogens and disease · 2022Article
Corrections and comments
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Authors and funding
2 authors.
Funding
Abstract
Chlamydia trachomatis is an obligate intracellular pathogen that has been historically difficult to genetically manipulate. Definitive progress in elucidating the mechanisms that C. trachomatis use to create and maintain a privileged intracellular niche has been limited due to a lack of genetic tools. Fortunately, there have recently been several new advances in genetic manipulation techniques. Among these is the development of fluorescence-reported allelic exchange mutagenesis (FRAEM). This method allows targeted gene deletion coupled with insertion of a selection cassette encoding antibiotic resistance and green fluorescent protein (GFP). Reliance on this strategy can be complicated when targeting genes within polycistronic operons due to the potential of polar effects on downstream genes. Floxed cassette allelic exchange mutagenesis (FLAEM), the protocol for which is described here, was developed to alleviate cassette-induced polar effects. FLAEM utilizes Cre-loxP genome editing to remove the selection cassette after targeted deletion by allelic exchange. The resulting strains contain markerless gene deletions of one or more coding sequences. This technique facilitates direct assessment of gene function and expands the repertoire of tools for genetic manipulation in C. trachomatis.
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Registered trials
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