| First Author | Canela A | Year | 2016 |
| Journal | Mol Cell | Volume | 63 |
| Issue | 5 | Pages | 898-911 |
| PubMed ID | 27477910 | Mgi Jnum | J:252872 |
| Mgi Id | MGI:6094124 | Doi | 10.1016/j.molcel.2016.06.034 |
| Citation | Canela A, et al. (2016) DNA Breaks and End Resection Measured Genome-wide by End Sequencing. Mol Cell 63(5):898-911 |
| abstractText | DNA double-strand breaks (DSBs) arise during physiological transcription, DNA replication, and antigen receptor diversification. Mistargeting or misprocessing of DSBs can result in pathological structural variation and mutation. Here we describe a sensitive method (END-seq) to monitor DNA end resection and DSBs genome-wide at base-pair resolution in vivo. We utilized END-seq to determine the frequency and spectrum of restriction-enzyme-, zinc-finger-nuclease-, and RAG-induced DSBs. Beyond sequence preference, chromatin features dictate the repertoire of these genome-modifying enzymes. END-seq can detect at least one DSB per cell among 10,000 cells not harboring DSBs, and we estimate that up to one out of 60 cells contains off-target RAG cleavage. In addition to site-specific cleavage, we detect DSBs distributed over extended regions during immunoglobulin class-switch recombination. Thus, END-seq provides a snapshot of DNA ends genome-wide, which can be utilized for understanding genome-editing specificities and the influence of chromatin on DSB pathway choice. |
