Mutagenic loads in cells edited through homologous recombination versus in trans paired nicking

Assessing the integrity of on-target and off-target genomic DNA sequences in cell populations subjected to canonical HR using regular or high-specificity nucleases or via in trans paired nicking using their corresponding D10A nickase derivatives. Targeted amplicon deep sequencing confirmed high and low indel frequencies at target sequences in cells exposed to nuclease and nickase complexes, respectively. Significantly, none of the nickase complexes tested induced detectable off-target activities. These experiments, strengthen the value of in trans paired nicking genome editing for targeted and precise chromosomal insertion of large genetic payloads with minimal bystander effects in genome-edited cell populations.