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Unlike CRISPR\u2013Cas9 systems that induce double-strand breaks, ABEs operate through precise deamination, avoiding chromosomal instability. However, the off-target editing effects of ABEs remain inadequately characterized. In this study, we present a biochemical method\u00a0Selict-seq, designed to evaluate genome-wide off-target editing by ABEs. Selict-seq specifically captures deoxyinosine-containing single-stranded DNA\u00a0and precisely identifies deoxyadenosine-to-deoxyinosine (dA-to-dI) mutation sites, elucidating the off-target effects induced by ABEs. Through investigations involving three single-guide RNAs, we identified numerous unexpected off-target edits both within and outside the protospacer regions. Notably, ABE8e(V106W) exhibited distinct off-target characteristics, including high editing rates (&amp;gt;10%) at previously unreported sites (e.g. RNF2 and EMX1) and out-of-protospacer mutations. These findings significantly advance our understanding of the off-target landscape associated with ABEs. In summary, our approach enables an unbiased analysis of the ABE editome and provides a widely applicable tool for specificity evaluation of various emerging genome editing technologies that produce intermediate products as deoxyinosine.<\/jats:p>","DOI":"10.1093\/nar\/gkaf281","type":"journal-article","created":{"date-parts":[[2025,3,31]],"date-time":"2025-03-31T14:20:37Z","timestamp":1743430837000},"source":"Crossref","is-referenced-by-count":17,"title":["Selict-seq profiles genome-wide off-target effects in adenosine base editing"],"prefix":"10.1093","volume":"53","author":[{"given":"Kexin","family":"Yuan","sequence":"first","affiliation":[{"name":"College of Chemistry and Molecular Sciences, Wuhan University , Wuhan , Hubei\u00a0 430072 ,","place":["PR China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Xi","sequence":"additional","affiliation":[{"name":"College of Chemistry and Molecular Sciences, Wuhan University , Wuhan , Hubei\u00a0 430072 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