Efficient Gene Editing with CRISPR RNPs

Cells were detached using Accutase, spun down at 600 g for 3 min and washed with PBS. Nucleofection was conducted using an Amaxa 96-well Shuttle system following the manufacturer’s protocol using 10 μl of Cas9 RNPs (100 pmole of Cas9, 120 pmole of guide RNAs) or Cpf1 RNPs (100 pmole of Cpf1, 120 pmole of crRNAs)^{49 (link)}. Cells (10⁵) were transfected using the EH-100 Lonza program. After nucleofection, medium (500 μl) was added and the cells were incubated at 37 °C in tissue culture plates. The cell culture medium was changed the next day, and the cells were then incubated for 7 days before flow cytometry analysis.