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DNA Breaks and End Resection Measured Genome-wide by End Sequencing.

Abstract：

: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.

journal_name

Mol Cell

journal_title

Molecular cell

authors

Canela A,Sridharan S,Sciascia N,Tubbs A,Meltzer P,Sleckman BP,Nussenzweig A

doi

10.1016/j.molcel.2016.06.034

subject

Has Abstract

pub_date

2016-09-01 00:00:00

pages

898-911

issue

5

eissn

1097-2765

issn

1097-4164

pii

S1097-2765(16)30292-1

journal_volume

63

pub_type

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