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| Main Authors: | , , , , , , |
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| Format: | Artículo científico |
| Language: | en |
| Published: |
Molecular plant-microbe interactions : MPMI
2025
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| Subjects: | |
| Online Access: | https://pubmed.ncbi.nlm.nih.gov/40752020/ |
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| _version_ | 1868266169510658050 |
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| author | Camborde, Laurent Jaturapaktrarak, Chalisa Gouzy, Jérôme Lopez-Roques, Céline Krajaejun, Theerapong Gaulin, Elodie Badis, Yacine |
| author_facet | Camborde, Laurent Jaturapaktrarak, Chalisa Gouzy, Jérôme Lopez-Roques, Céline Krajaejun, Theerapong Gaulin, Elodie Badis, Yacine Camborde, Laurent Jaturapaktrarak, Chalisa Gouzy, Jérôme Lopez-Roques, Céline Krajaejun, Theerapong Gaulin, Elodie Badis, Yacine |
| collection | PubMed - marine biology |
| contents | Adenine Phosphoribosyltransferase Is a Universal Counterselectable Marker for DNA-Free Genome Editing in Oomycetes. Camborde, Laurent Jaturapaktrarak, Chalisa Gouzy, Jérôme Lopez-Roques, Céline Krajaejun, Theerapong Gaulin, Elodie Badis, Yacine Gene Editing CRISPR-Cas Systems Adenine Phosphoribosyltransferase Oomycetes Phytophthora Mutation CRISPR-Cas genome editing is a powerful tool for understanding the pathogenicity of oomycetes, a group that includes several destructive plant parasites. Although a few species have benefited from plasmid-based transformation methods for gene overexpression and RNA interference silencing, these techniques remain inefficient for other oomycete genera such as and . Here, we explored the applicability of DNA-free endogenous counterselection in filamentous oomycetes, using CRISPR-Cas9 ribonucleoproteins (RNPs). We used biolistics to deliver RNPs targeting the adenine phosphoribosyltransferase () gene and generated selectable 2-fluoroadenine-resistant mutants in , , and species. Targeted mutagenesis resulted in various deletions at the expected cut sites, confirming efficient genome editing. Knockout mutants exhibited no alterations in growth or virulence, making a suitable selectable marker gene for oomycete research. Whole-genome comparative analyses on CRISPR-edited mutants revealed no or very few additional mutations in . and and substantial off-target effects in This one-step approach circumvents the need for protoplast generation and can be broadly applied to oomycetes producing zoospores or oospores. [Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license. |
| format | Artículo científico |
| id | pubmed_40752020 |
| institution | PubMed |
| language | en |
| publishDate | 2025 |
| publisher | Molecular plant-microbe interactions : MPMI |
| record_format | pubmed |
| spellingShingle | Adenine Phosphoribosyltransferase Is a Universal Counterselectable Marker for DNA-Free Genome Editing in Oomycetes. Camborde, Laurent Jaturapaktrarak, Chalisa Gouzy, Jérôme Lopez-Roques, Céline Krajaejun, Theerapong Gaulin, Elodie Badis, Yacine Gene Editing CRISPR-Cas Systems Adenine Phosphoribosyltransferase Oomycetes Phytophthora Mutation Adenine Phosphoribosyltransferase Is a Universal Counterselectable Marker for DNA-Free Genome Editing in Oomycetes. Camborde, Laurent Jaturapaktrarak, Chalisa Gouzy, Jérôme Lopez-Roques, Céline Krajaejun, Theerapong Gaulin, Elodie Badis, Yacine Gene Editing CRISPR-Cas Systems Adenine Phosphoribosyltransferase Oomycetes Phytophthora Mutation CRISPR-Cas genome editing is a powerful tool for understanding the pathogenicity of oomycetes, a group that includes several destructive plant parasites. Although a few species have benefited from plasmid-based transformation methods for gene overexpression and RNA interference silencing, these techniques remain inefficient for other oomycete genera such as and . Here, we explored the applicability of DNA-free endogenous counterselection in filamentous oomycetes, using CRISPR-Cas9 ribonucleoproteins (RNPs). We used biolistics to deliver RNPs targeting the adenine phosphoribosyltransferase () gene and generated selectable 2-fluoroadenine-resistant mutants in , , and species. Targeted mutagenesis resulted in various deletions at the expected cut sites, confirming efficient genome editing. Knockout mutants exhibited no alterations in growth or virulence, making a suitable selectable marker gene for oomycete research. Whole-genome comparative analyses on CRISPR-edited mutants revealed no or very few additional mutations in . and and substantial off-target effects in This one-step approach circumvents the need for protoplast generation and can be broadly applied to oomycetes producing zoospores or oospores. [Formula: see text] Copyright © 2025 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license. |
| title | Adenine Phosphoribosyltransferase Is a Universal Counterselectable Marker for DNA-Free Genome Editing in Oomycetes. |
| topic | Gene Editing CRISPR-Cas Systems Adenine Phosphoribosyltransferase Oomycetes Phytophthora Mutation |
| url | https://pubmed.ncbi.nlm.nih.gov/40752020/ |