Identification of DNA Single-strand Breaks with Single Nucleotide Resolution
Defective repair of DNA single-strand breaks (SSBs) is implicated in neurodegenerative disorders and cancer. Furthermore, recent advances in genome editing rely on nicking DNA at specific loci and may be susceptible to generating off-target SSBs. Despite this, current methods for measuring such breaks are lacking. Here, we present NickSeq, a method that is uniquely capable of identifying SSBs with strand-specific single nucleotide resolution in an unbiased manner. We then utilize this method to identify the off-target activity of Cas9 nickase.