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Related Experiment Video

Updated: Feb 5, 2026

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira

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Published on: August 14, 2021

Generating Gene Silenced Mutants in Phytophthora sojae.

Yao Zhao^1,2, Yufei Li¹, Min Qiu¹

¹Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.

Methods in Molecular Biology (Clifton, N.J.)

|September 6, 2018

Summary

Gene silencing in Phytophthora sojae offers a valuable alternative to gene knockout for studying gene function, especially when lethality is a concern. This method uses antisense RNA to reduce target gene expression in the soybean pathogen.

Keywords:

Antisense RNA Gene silencing PEG-mediated transformation

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Area of Science:

Plant Pathology
Molecular Biology
Agricultural Science

Background:

Phytophthora species are significant plant pathogens causing substantial agricultural losses.
Understanding Phytophthora pathogenesis requires effective gene function investigation methods.
CRISPR/Cas9 gene knockout has limitations, particularly when mutations are lethal.

Purpose of the Study:

To introduce a gene silencing protocol for the soybean pathogen, Phytophthora sojae.
To provide a viable alternative to gene knockout for functional gene analysis in Phytophthora.

Main Methods:

Development of a gene silencing protocol for Phytophthora sojae.
Utilizing PEG-mediated protoplast transformation to integrate an expression cassette.
Constitutive production of antisense RNA transcripts targeting specific mRNAs.

Main Results:

Successful implementation of a gene silencing strategy in P. sojae.
Antisense RNA transcripts effectively target complementary mRNAs.
Significant reduction in the expression of target genes achieved.

Conclusions:

Gene silencing is a crucial tool for functional gene analysis in Phytophthora, complementing gene knockout methods.
The described protocol enables effective gene silencing in the soybean pathogen P. sojae.
This approach facilitates deeper understanding of Phytophthora pathogenesis.