Three delivery methods of CRISPR/Cas9 to induce targeted mutations into centromeric histone H3 (CENH3) in carrots (Daucus carota L.)

Key Points

• This study aims to evaluate different CRISPR/Cas9 delivery methods to mutate CENH3 in carrots.
• Used CRISPR/Cas9 to target CENH3 in carrot transformations.
• Conducted transformations using Agrobacterium tumefaciens and Agrobacterium rhizogenes.
• Employed transient transformation of protoplasts with CRISPR/Cas9 RNPs.
• Transformation with Agrobacterium led to secondary mutations and lower CENH3 levels.
• Protoplasts transformed with RNPs were transgene-free and successfully mutated.
• The transgene-free method was the most effective, minimizing secondary mutations.