PT  - JOURNAL ARTICLE
AU  - Beeke Wienert
AU  - Stacia K Wyman
AU  - Christopher D Richardson
AU  - Charles D Yeh
AU  - Pinar Akcakaya
AU  - Michelle J Porritt
AU  - Michaela Morlock
AU  - Jonathan T Vu
AU  - Katelynn R Kazane
AU  - Hannah L Watry
AU  - Luke M Judge
AU  - Bruce R Conklin
AU  - Marcello Maresca
AU  - Jacob E Corn
TI  - Unbiased detection of CRISPR off-targets <em>in vivo</em> using DISCOVER-Seq
AID  - 10.1101/469635
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 469635
4099  - http://biorxiv.org/content/early/2018/11/14/469635.short
4100  - http://biorxiv.org/content/early/2018/11/14/469635.full
AB  - Genome editing using nucleases such as CRISPR-Cas induces programmable DNA damage at a target genomic site but can also affect off-target sites. Here, we develop a powerful, sensitive assay for the unbiased identification of off-target sites that we term DISCOVER-Seq. This approach takes advantage of the recruitment of endogenous DNA repair factors for genome-wide identification of Cas-induced double-strand breaks. One such factor, MRE11, is recruited precisely to double-strand breaks, enabling molecular characterization of nuclease cut sites with single-base resolution. DISCOVER-Seq detects off-targets in cellular models and in vivo upon adenoviral gene editing of mouse livers, paving the way for real-time off-target discovery during therapeutic gene editing. DISCOVER-Seq is furthermore applicable to multiple types of Cas nucleases and provides an unprecedented view of events that precede repair of the affected sites.