CRISPR/Cas9-induced site-specific DNA double-strand breaks (DSBs) can be repaired by homology-directed repair (HDR) or non-homologous end joining (NHEJ) pathways.Extensive efforts have been made to knock-in exogenous DNA to a selected genomic locus in human cells;which,however,has focused on HDR-based strategies andwas proven inefficient.Here,we report that NHEJ pathway mediates efficient rejoining of genome and plasmids following CRISPR/Cas9-induced DNA DSBs,and promotes high-efficiencyDNA integration in various human cell types.