An efficient CRISPR/Cas9 platform for targeted genome editing in rose (Rosa hybrida)

Area of Science:

• Plant Biotechnology
• Molecular Biology
• Genomics

Background:

• The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-related nuclease 9 (Cas9) system is a powerful gene editing tool, but its application in roses (Rosa hybrida) has been limited.
• Roses possess complex genomes and lack efficient transformation methods, hindering the adoption of CRISPR/Cas9 technology.

Purpose of the Study:

• To establish an efficient CRISPR/Cas9 gene editing platform for roses.
• To optimize single-guide RNA (sgRNA) screening for high editing efficiency in rose suspension cells.
• To demonstrate the system's utility by editing the RhEIN2 gene and inducing ethylene insensitivity.

Main Methods:

• Developed a suspension cell-based platform for screening highly efficient sgRNAs for CRISPR/Cas9 in roses.
• Utilized the Arabidopsis thaliana U6-29 promoter to drive sgRNA expression, enhancing the CRISPR/Cas9 system's activity.
• Applied the optimized CRISPR/Cas9 system to target and edit the RhEIN2 gene in roses.

Main Results:

• Successfully established an efficient CRISPR/Cas9 gene editing system for roses.
• Identified highly effective sgRNAs for precise gene editing in rose suspension cells.
• Achieved successful editing of the RhEIN2 gene, resulting in ethylene insensitivity in the modified roses.

Conclusions:

• The optimized CRISPR/Cas9 system offers a robust tool for gene editing in roses.
• This technology facilitates functional genomics, molecular breeding, and synthetic biology applications in rose research.
• The development opens new avenues for genetic improvement and trait manipulation in roses.