Sensitive and unbiased genome-wide profiling of base-editor-induced off-target activity using CHANGE-seq-BE

Detection of the off-target effects of base editors is important for identifying their safety risks but current methods for understanding their global activities have limitations in terms of sensitivity or bias by computationally selecting a subset of sites for experimental analysis. We present CHANGE-seq-BE, a method to assess the guide RNA-dependent off-target profile of both adenine and cytosine base editors that is simultaneously sensitive and unbiased. CHANGE-seq-BE relies on selective sequencing of base-editor-modified genomic DNA in vitro and provides comprehensive identification of genome-wide off-target mutations. We found that 98.8% of validated off-target sites were unique to ABE8e adenine base…

• DD
  Doris Duke Charitable Foundation

  Award: 2020154
• NH
  National Heart, Lung, and Blood Institute

  Award: U01HL163983
• NI
  National Institute of Allergy and Infectious Diseases

  Awards: U01AI176471, U01AI176470