Editing efficiencies of Cas9 and highly stable Cas9 on HBV genome
HuH7 cells were seeded into 3 wells of 6-well plate and co-transfected with pBB4.5-1.3HBV, PX458-Cas9 or PX458-HSCas9, and pU6 plasmids (NC) or pU6-t-g1-t-g2-t plasmids, respectively. At 4 days post of transfection, the cells were collected for extracting total DNA, followed by PCR with the primers outsides two cleave sites and next-generation sequencing assay. Editing efficiency was the number of non-wild-type reads at the cleavage site divided by the total number of reads at the cleavage site.