Engineering Citrobacter freundii using CRISPR/Cas9 system Article Swipe
Trinidad Alfaro
,
Joshua R. Elmore
,
Zachary R. Stromberg
,
Janine R. Hutchison
,
Becky Hess
·
YOU?
·
· 2022
· Open Access
·
· DOI: https://doi.org/10.1016/j.mimet.2022.106533
YOU?
·
· 2022
· Open Access
·
· DOI: https://doi.org/10.1016/j.mimet.2022.106533
The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR associated proteins) system is a useful tool to edit genomes quickly and efficiently. However, the use of CRISPR/Cas9 to edit bacterial genomes has been limited to select microbial chassis primarily used for bioproduction of high value products. Thus, expansion of CRISPR/Cas9 tools to other microbial organisms is needed. Here, our aim was to assess the suitability of CRISPR/Cas9 for genome editing of the Citrobacter freundii type strain ATCC 8090. We evaluated the commonly used two plasmid pCas/pTargetF system to enable gene deletions and insertions in C. freundii and determined editing efficiency. The CRISPR/Cas9 based method enabled high editing efficiency (~91%) for deletion of galactokinase (galk) and enabled deletion with various single guide RNA (sgRNA) sequences. To assess the ability of CRISPR/Cas9 tools to insert genes, we used the fluorescent reporter mNeonGreen, an endopeptidase (yebA), and a transcriptional regulator (xylS) and found successful insertion with high efficiency (81-100%) of each gene individually. These results strengthen and expand the use of CRISPR/Cas9 genome editing to C. freundii as an additional microbial chassis.
Related Topics
Concepts
CRISPR
Genome editing
Citrobacter freundii
Cas9
Biology
Trans-activating crRNA
Plasmid
Computational biology
Gene
Genetics
CRISPR interference
Enterobacteriaceae
Escherichia coli
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1016/j.mimet.2022.106533
- OA Status
- hybrid
- Cited By
- 4
- References
- 36
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4283728314
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4283728314Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1016/j.mimet.2022.106533Digital Object Identifier
- Title
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Engineering Citrobacter freundii using CRISPR/Cas9 systemWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2022Year of publication
- Publication date
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2022-06-29Full publication date if available
- Authors
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Trinidad Alfaro, Joshua R. Elmore, Zachary R. Stromberg, Janine R. Hutchison, Becky HessList of authors in order
- Landing page
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https://doi.org/10.1016/j.mimet.2022.106533Publisher landing page
- Open access
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YesWhether a free full text is available
- OA status
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hybridOpen access status per OpenAlex
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https://doi.org/10.1016/j.mimet.2022.106533Direct OA link when available
- Concepts
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CRISPR, Genome editing, Citrobacter freundii, Cas9, Biology, Trans-activating crRNA, Plasmid, Computational biology, Gene, Genetics, CRISPR interference, Enterobacteriaceae, Escherichia coliTop concepts (fields/topics) attached by OpenAlex
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4Total citation count in OpenAlex
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2024: 1, 2023: 2, 2022: 1Per-year citation counts (last 5 years)
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36Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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