JC
Jingting Cao
  • Department of Plant Pathology, Nanjing Agricultural University, Nanjing, Jiangsu 210095
Phytophthora sojae Transformation Based on the CRISPR/Cas9 System
Authors:  Jingting Cao, Min Qiu, Wenwu Ye and Yuanchao Wang, date: 03/20/2022, view: 2124, Q&A: 0

Phytophthora sojae is a model species for the study of plant pathogenic oomycetes. The initial research on gene function using Phytophthora was mainly based on gene silencing technology. Recently, the CRISPR/Cas9-mediated genome editing technology was successfully established in P. sojae and widely used in oomycetes. In this protocol, we describe the operating procedures for the use of CRISPR/Cas9-based genome editing technology and PEG-mediated stable transformation of P. sojae protoplasts. Two plasmids were co-transformed into P. sojae: pYF515 expressing Cas9 and the single guide RNA, and the homologous replacement vector of the candidate gene. Finally, the ORF of candidate gene were replaced with the ORF of the entire hygromycin B phosphotransferase gene (HPH), to achieve precise knockout.

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