iScienceSep 8, 2026

A simple method for making fungal cells and editing their genes using CRISPR/Cpf1 in Fusarium oxysporum

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Sep 9, 2026

Abstract

An efficient CRISPR/Cpf1 system was engineered to improve gene-editing methods for Fusarium oxysporum.

Innovations include small-scale protoplast generation that reduces enzyme use and simplifies workflows.
The CRISPR/Cpf1 system allows for shorter guide RNA design and staggered DNA cleavage to enhance gene targeting.
Minimal homology arm strategies significantly lessen cloning complexity.
Extensive validation confirms successful gene targeting, with verification and functional analysis using standardized pathogenicity assays.
The platform provides affordable tools for advancing research on F. oxysporum, potentially benefiting agricultural biotechnology.

Simplified

Fusarium oxysporum is a significant threat to agriculture and One Health, requiring advanced molecular tools for functional genomic analyses and biological control agent development. Existing gene-editing methods are hampered by costly protoplast preparation protocols and by CRISPR-Cas9 limitations, such as restricted protospacer adjacent motif (PAM) sequences and complex guide RNA requirements. We engineered an efficient CRISPR/Cpf1 system that overcomes these issues through three main innovations: small-scale protoplast generation using filter column-based methods that greatly reduce enzyme consumption while simplifying workflows, a CRISPR/Cpf1 system with shorter guide RNA design and staggered DNA cleavage to promote homologous recombination, and minimal homology arm strategies that significantly decrease cloning complexity. Extensive validation confirms successful gene targeting with molecular verification and functional analysis via standardized pathogenicity assays. This integrated platform offers affordable, accessible tools for systematic F. oxysporum research, enhancing fundamental understanding of plant-pathogen interactions and supporting high-throughput screening vital for agricultural biotechnology and biological agent development.

Full Text

Full text is available at the source.

View Full Text ↗

A simple method for making fungal cells and editing their genes using CRISPR/Cpf1 in Fusarium oxysporum

Abstract

Full Text

You found one interesting study. We’ll send the next 7.

What Lands in Your Inbox Each Week:

Recent issues from the crispr gene editing brief

Abstract

Full Text

Related papers

You found one interesting study. We’ll send the next 7.

What Lands in Your Inbox Each Week:

Recent issues from the crispr gene editing brief