On-target and Off-target Activities of RAG Are Restricted by Topological Chromatin Domains

ABSTRACT: RAG initiates V(D)J recombination in developing lymphocytes by generating "on-target" DNA double-stranded breaks at bona fide recombination signal sequence (RSS) pairs. We now employ RAG-generated DNA breaks in endogenous or ectopically-inserted RSS pairs as bait to identify huge numbers of RAG "off-target" sites. These off-target DNA breaks occur across convergent CTCF-binding element (CBE)-flanked loop domains containing the bait RSS pairs. Such off-target cleavage occurs at the simple CAC motif that defines the RSS cleavage-site and shows orientation-dependence best explained by a two-dimensional tracking mechanism. Deletion of the CBE-based IGCR1 IgH regulatory element disrupts antibody IgH recombination domains and, correspondingly, alters distributions of RAG on

ORGANISM(S): Mus musculus

SUBMITTER: Jiazhi Hu 

PROVIDER: E-GEOD-73007 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress