Designing CRISPR/Cas9 System Targeting OsERF922 Gene of J02 Rice Variety

Abstract

Background and Objective: AP2/ERF has been known as a key transcription factor that involves biotic stress responses in rice (Oryza sativa). However, little information is available on the involvement of AP2/ERF in response to rice blast disease. In this study, the expression level of the rice OsERF922 gene that encodes AP2/ERF was analyzed in the J02 variety treated with Magnaporthe oryzae, the causal agent of rice blast disease and a CRISPR/Cas9 targeting the OsERF922 was engineered for further study the functions of OsERF922. Materials and Methods: The expression of OsERF922 in rice was measured by Reverse Transcription PCR (RT-PCR). The Single Guide RNAs (sgRNAs) targeting OsERF922 were designed by CRISPR-P 2.0 software and constructed into a pENTR4 vector. Results: RT-PCR analysis revealed that the expression of OsERF922 of the treated plants was up-regulated compared to those in controls. In addition, two sgRNA sequences targeting OsERF922 were successfully cloned into the expression vector. Conclusion: The results provided useful materials for further functional characterization of OsERF922 by the CRISPR/Cas9 system.