A Cas12a‐based gene editing system for <i>Phytophthora infestans</i> reveals monoallelic expression of an elicitor

Ah‐Fong, Audrey M. V.; Boyd, Amy M.; Matson, Michael Edmund Hans; Judelson, Howard S.

doi:10.1111/mpp.13051

Cited by 29 publications

(37 citation statements)

References 83 publications

(121 reference statements)

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“…The application of CRISPR/Cas has addressed many of the previous obstacles for editing oomycete genes, and successful transformants have been generated, such as from the use of a CRISPR/Cas12a system in P. infestans, which has enabled a greater capability for gene editing in oomycetes generally [64]. Similarly, point mutations introduced into P. sojae [65] using CRISPR/Cas9 conferred resistance to the fungicide oxathiapiprolin.…”

Section: Gene Editing Using Crispr/casmentioning

confidence: 99%

How to Unravel the Key Functions of Cryptic Oomycete Elicitin Proteins and Their Role in Plant Disease

Kharel

Islam

Rookes

et al. 2021

Plants

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Pathogens and plants are in a constant battle with one another, the result of which is either the restriction of pathogen growth via constitutive or induced plant defense responses or the pathogen colonization of plant cells and tissues that cause disease. Elicitins are a group of highly conserved proteins produced by certain oomycete species, and their sterol binding ability is recognized as an important feature in sterol–auxotrophic oomycetes. Elicitins also orchestrate other aspects of the interactions of oomycetes with their plant hosts. The function of elicitins as avirulence or virulence factors is controversial and is dependent on the host species, and despite several decades of research, the function of these proteins remains elusive. We summarize here our current understanding of elicitins as either defense-promoting or defense-suppressing agents and propose that more recent approaches such as the use of ‘omics’ and gene editing can be used to unravel the role of elicitins in host–pathogen interactions. A better understanding of the role of elicitins is required and deciphering their role in host–pathogen interactions will expand the strategies that can be adopted to improve disease resistance and reduce crop losses.

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Section: Gene Editing Using Crispr/casmentioning

confidence: 99%

How to Unravel the Key Functions of Cryptic Oomycete Elicitin Proteins and Their Role in Plant Disease

Kharel

Islam

Rookes

et al. 2021

Plants

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“…Meanwhile, some studies have shown that the off-target effect is extremely low in iPSCs [44,45]. Compared with Cas9, Cas12a is more sensitive to mismatches in the guide RNA and has lower off-target effects [46]. In this study, no off-target cleavage was observed at the seven putative sites, even when the COE expression lasted 75 days.…”

Section: Discussionmentioning

confidence: 51%

An Episomal CRISPR/Cas12a System for Mediating Efficient Gene Editing

Duan

Tang

Zeng

et al. 2021

Life

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(1) Background: Gene editing technology, as represented by CRISPR is a powerful tool used in biomedical science. However, the editing efficiency of such technologies, especially in induced pluripotent stem cells (iPSCs) and other types of stem cells, is low which hinders its application in regenerative medicine; (2) Methods: A gene-editing system, COE, was designed and constructed based on CRISPR/Cas12a and Orip/EBNA1, and its editing efficiency was evaluated in human embryonic kidney 293T (HEK-293T) cells with flow cytometry and restriction fragment length polymorphism (RFLP) analysis. The COE was nucleofected into iPSCs, then, the editing efficiency was verified by a polymerase chain reaction and Sanger sequencing; (3) Results: With the extension of time, COE enables the generation of up to 90% insertion or deletion rates in HEK-293T cells. Furthermore, the deletion of a 2.5 kb fragment containing Exon 51 of the dystrophin gene (DMD) in iPSCs was achieved with high efficiency; out of 14 clones analyzed, 3 were positive. Additionally, the Exon 51-deleted iPSCs derived from cardiomyocytes had similar expression profiles to those of Duchenne muscular dystrophy (DMD) patient-specific iPSCs. Moreover, there was no residue of each component of the plasmid in the editing cells; (4) Conclusions: In this study, a novel, efficient, and safe gene-editing system, COE, was developed, providing a powerful tool for gene editing.

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“…In P. infestans , however, CRISPR-Cas9-mediated gene editing was not successful ( van den Hoogen and Govers, 2018b ). A recent study by Ah-Fong et al (2021) showed that Cas9 is toxic for this species, but with Cas12a as nuclease they obtained transformants that are viable and have small deletions in the target gene, the inf1 elicitin gene. This is another major leap forward as it demonstrates the successful implementation of a promising gene editing tool in P. infestans .…”

Section: The Interplay Between Host and Oomycete Pathogensmentioning

confidence: 99%

Uncovering the Role of Metabolism in Oomycete–Host Interactions Using Genome-Scale Metabolic Models

et al. 2021

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Metabolism is the set of biochemical reactions of an organism that enables it to assimilate nutrients from its environment and to generate building blocks for growth and proliferation. It forms a complex network that is intertwined with the many molecular and cellular processes that take place within cells. Systems biology aims to capture the complexity of cells, organisms, or communities by reconstructing models based on information gathered by high-throughput analyses (omics data) and prior knowledge. One type of model is a genome-scale metabolic model (GEM) that allows studying the distributions of metabolic fluxes, i.e., the “mass-flow” through the network of biochemical reactions. GEMs are nowadays widely applied and have been reconstructed for various microbial pathogens, either in a free-living state or in interaction with their hosts, with the aim to gain insight into mechanisms of pathogenicity. In this review, we first introduce the principles of systems biology and GEMs. We then describe how metabolic modeling can contribute to unraveling microbial pathogenesis and host–pathogen interactions, with a specific focus on oomycete plant pathogens and in particular Phytophthora infestans. Subsequently, we review achievements obtained so far and identify and discuss potential pitfalls of current models. Finally, we propose a workflow for reconstructing high-quality GEMs and elaborate on the resources needed to advance a system biology approach aimed at untangling the intimate interactions between plants and pathogens.

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A Cas12a‐based gene editing system for Phytophthora infestans reveals monoallelic expression of an elicitor

Cited by 29 publications

References 83 publications

How to Unravel the Key Functions of Cryptic Oomycete Elicitin Proteins and Their Role in Plant Disease

How to Unravel the Key Functions of Cryptic Oomycete Elicitin Proteins and Their Role in Plant Disease

An Episomal CRISPR/Cas12a System for Mediating Efficient Gene Editing

Uncovering the Role of Metabolism in Oomycete–Host Interactions Using Genome-Scale Metabolic Models

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