Gene Editing and Protein Tagging in the Oomycete Phytophthora infestans Using CRISPR-Cas12a.

Carl S Mendoza, Audrey M V Ah-Fong, Howard S Judelson

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Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

3 authors.

Carl S MendozaDepartment of Microbiology and Plant Pathology, University of California, Riverside, CA, USA.

Audrey M V Ah-FongDepartment of Microbiology and Plant Pathology, University of California, Riverside, CA, USA.

Howard S JudelsonDepartment of Microbiology and Plant Pathology, University of California, Riverside, CA, USA. judelson@ucr.edu.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Molecular genetic tools such as CRISPR-Cas gene editing systems are invaluable for understanding gene and protein function and revealing the details of a pathogen's life and disease cycles. Here we present protocols for genome editing in Phytophthora infestans, an oomycete with global importance as a pathogen of potato and tomato. Using a vector system that expresses variants of Cas12a from Lachnospiraceae bacterium and its guide RNA from a unified transcript, we first present a method for editing genes through the non-homologous end-joining (NHEJ) pathway. We then describe an application of homology-directed repair (HDR), in which Cas12a is used to fuse a protein-coding gene with a fluorescent or epitope tag. Both methods should be adaptable to many oomycetes other than P. infestans.

Indexed as

CRISPR-Cas SystemsGene EditingPhytophthora infestansBacterial ProteinsCRISPR-Associated ProteinsDNA End-Joining RepairEndodeoxyribonucleasesGenetic VectorsRecombinational DNA RepairRNA, Guide, CRISPR-Cas SystemsBacterial ProteinsCas12a proteinCRISPR-Associated ProteinsEndodeoxyribonucleasesRNA, Guide, CRISPR-Cas SystemsCRISPRFluorescent reporterGene editingOomyceteTransformation

Identifiers

PMID39729268

DOI10.1016/j.molcel.2018.06.043

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.