Figure 3: Flowchart of CRISPR/Cas9 RNP induced targeted mutagenesis in wheat.
From: Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes
RNP preparation takes 3–4 days (including Cas9 protein purification). RNP validation in vitro and in protoplasts takes 3–4 days (not including the time for plant preparation). RNP coating and delivery can be completed within 4 h. After this, the plantlets regenerate in 6–8 weeks. Identification of mutants by PCR-RE and DNA sequencing takes 3 days.
