design: The GUIDE-seq experiments were performed in human HEK293T and mouse Neuro-2a cells. Generally, A total of 3  105 HEK293T or Neuro-2a cells were transfected with Cas9/sgRNA ribonucleoprotein (RNP), together with 1.5 ug annealed double-stranded oligodeoxynucleotide by electroporation. After 48-72 hours culture, cells were collected for genomic DNA extraction with the TIANamp Genomic DNA Kit (TIANGEN) for GUIDE-seq analysis (GENEWIZ, Azenta Life Sciences).