New Publication: Dual‑gRNA CRISPR/Cas9 Deletion of CsDMR6 in Sweet Orange

This week, our team’s latest research has been published in Plants (MDPI), presenting a significant advance in the genetic improvement of citrus crops. The study demonstrates the successful deletion of the CsDMR6 gene in Citrus sinensis using a dual‑gRNA CRISPR/Cas9 system, supported by an optimized in vitro regeneration protocol for juvenile sweet orange genotypes.

Huanglongbing (HLB) remains the most destructive disease affecting citrus worldwide. By targeting CsDMR6—a susceptibility‑associated gene—our work provides a promising route toward developing HLB‑tolerant citrus plants through precise genome editing.

Key Highlights of the Study

• Development of improved regeneration conditions using Citrus Shoot Multiplication (CSM) and Citrus Callus Induction (CIM) media for ‘Valencia’ sweet orange and hybrid lines CsH1–CsH3.

• Enhanced morphogenic responses achieved through the use of GA₃ and NAA.

• Transient validation of CRISPR activity via GFP fluorescence, diagnostic PCR (~447 bp), and Sanger sequencing.

• Efficient large‑fragment deletions confirmed between two CRISPR target sites, demonstrating the robustness of the dual‑gRNA strategy.

This publication marks an important step toward applying advanced gene‑editing tools to strengthen citrus resilience against HLB. It also reinforces our team’s leadership in plant biotechnology and the development of innovative solutions for high‑impact agricultural challenges.