ArticleMolecular plant pathology2021
A Cas12a-based gene editing system for Phytophthora infestans reveals monoallelic expression of an elicitor.
Article in Molecular plant pathology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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20 citing papers in PubMed, 36 citations in OpenAlex.
- Novel application of ribonucleoprotein-mediated CRISPR-Cas9 gene editing in plant pathogenic oomycete species.Microbiology spectrum · 2025Article
- Transient expression of fluorescent proteins and Cas nucleases inMicrobiology (Reading, England) · 2025Article
- Gene Editing and Protein Tagging in the Oomycete Phytophthora infestans Using CRISPR-Cas12a.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Transcription factor binding specificities of the oomycete Phytophthora infestans reflect conserved and divergent evolutionary patterns and predict function.BMC genomics · 2024Article
- Modern Breeding Strategies and Tools for Durable Late Blight Resistance in Potato.Plants (Basel, Switzerland) · 2024Review
- Review
- Heterogeneity in establishment of polyethylene glycol-mediated plasmid transformations for five forest pathogenic Phytophthora species.PloS one · 2024Article
- Breeding strategies for late blight resistance in potato crop: recent developments.Molecular biology reports · 2023Review
- A modified Agrobacterium-mediated transformation for two oomycete pathogens.PLoS pathogens · 2023Article
- Fungal biotechnology: From yesterday to tomorrow.Frontiers in fungal biology · 2023Review
- Progress and Challenges in Elucidating the Functional Role of Effectors in the Soybean-Journal of fungi (Basel, Switzerland) · 2022Review
- CRISPR-Cas12a induced DNA double-strand breaks are repaired by multiple pathways with different mutation profiles in Magnaporthe oryzae.Nature communications · 2022Article
- The contribution of DNA repair pathways to genome editing and evolution in filamentous pathogens.FEMS microbiology reviews · 2022Review
- Signal and regulatory mechanisms involved in spore development ofFrontiers in microbiology · 2022Review
- An Episomal CRISPR/Cas12a System for Mediating Efficient Gene Editing.Life (Basel, Switzerland) · 2021Article
- Specific interaction of an RNA-binding protein with the 3'-UTR of its target mRNA is critical to oomycete sexual reproduction.PLoS pathogens · 2021Article
- How to Unravel the Key Functions of Cryptic Oomycete Elicitin Proteins and Their Role in Plant Disease.Plants (Basel, Switzerland) · 2021Review
- A Cas12a-based gene editing system for Phytophthora infestans reveals monoallelic expression of an elicitor.Molecular plant pathology · 2021Article
- The Mevalonate Pathway Is Important for Growth, Spore Production, and the Virulence ofFrontiers in microbiology · 2021Article
- Uncovering the Role of Metabolism in Oomycete-Host Interactions Using Genome-Scale Metabolic Models.Frontiers in microbiology · 2021Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Phytophthora infestans is a destructive pathogen of potato and a model for investigations of oomycete biology. The successful application of a CRISPR gene editing system to P. infestans is so far unreported. We discovered that it is difficult to express CRISPR/Cas9 but not a catalytically inactive form in transformants, suggesting that the active nuclease is toxic. We were able to achieve editing with CRISPR/Cas12a using vectors in which the nuclease and its guide RNA were expressed from a single transcript. Using the elicitor gene Inf1 as a target, we observed editing of one or both alleles in up to 13% of transformants. Editing was more efficient when guide RNA processing relied on the Cas12a direct repeat instead of ribozyme sequences. INF1 protein was not made when both alleles were edited in the same transformant, but surprisingly also when only one allele was altered. We discovered that the isolate used for editing, 1306, exhibited monoallelic expression of Inf1 due to insertion of a copia-like element in the promoter of one allele. The element exhibits features of active retrotransposons, including a target site duplication, long terminal repeats, and an intact polyprotein reading frame. Editing occurred more often on the transcribed allele, presumably due to differences in chromatin structure. The Cas12a system not only provides a tool for modifying genes in P. infestans, but also for other members of the genus by expanding the number of editable sites. Our work also highlights a natural mechanism that remodels oomycete genomes.
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