Dna-free gene editing for early flowering in brassica napus

Overview

This innovative solution employs the CRISPR/Cas9 gene-editing technology to target and knockout key flowering repressors, FT and TFL1, in Brassica napus. By using protoplast-mediated transformation for DNA-free delivery of Cas9 Ribonucleoproteins (RNPs), the method ensures non-transgenic production. This results in a significantly shorter time to flowering, advancing the process by over 15-20 days, and allows for the regeneration of stable, edited lines without transgenes.

Technical specifications

• CRISPR/Cas9 Technology: Utilizes precise genome editing to target FT/TFL1 orthologs.
• Protoplast-Mediated Transformation: Offers a DNA-free delivery method for Cas9 RNPs, bypassing the integration of foreign DNA and ensuring non-transgenic plants.
• Multiplexed CRISPR System: Capable of simultaneously targeting up to eight gene copies in the complex Brassica napus genome.
• Validation Process: Includes cloning, PEG-mediated transformation, and PCR plus sequencing confirmation of knockout T0 plants.

Technology readiness level

The technology is at TRL 4, with successful proof-of-concept in Petunia and a structured plan for further validation in Brassica napus. The approach is supported by laboratory capabilities and expertise at AURAK, UAE, with a 36-month timeline for development and validation.