Comparative analysis of CRISPR/Cas9 delivery approaches for polyphenol oxidase 2 gene editing in potato

Abstract

StPPO2 gene editing was analyzed by Agrobacterium mediated transformation with CR-PPO vector, ribonucleoprotein complexes (RNP-PPO) transfection to protoplasts, and CR-PPO transient expression in protoplasts, yielding efficiencies of 9.6%, 18.4%, and 31.9%, respectively. Transient expression of CR-PPO in protoplasts resulted in tetra-allelic edited lines, observed in 46% of total edited lines. On-target DNA insertions were found in lines from all three approaches. Loss of function of the StPPO2 protein was confirmed in a tetra-allelic edited line.