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By John D. Curry, Danae Schulz, Cynthia J. Guidos, Jayne S. Danska, Lauryl Nutter, Andre Nussenzweig and Mark S. Schlissel


transferred-end capture assay; NHEJ, nonhomologous end joining; RSS, recombination signal sequence. The V(D)J recombinase catalyzes DNA transposition and translocation both in vitro and in vivo. Because lymphoid malignancies contain chromosomal translocations involving antigen receptor and protooncogene loci, it is critical to understand the types of “ mistakes ” made by the recombinase. Using a newly devised assay, we characterized 48 unique TCR � recombination signal sequence (RSS) end insertions in murine thymocyte and splenocyte genomic DNA samples. Nearly half of these events targeted “ cryptic ” RSS-like elements. In no instance did we detect target-site duplications, which is a hallmark of recombinase-mediated transposition in vitro. Rather, these insertions were most likely caused by either V(D)J recombination between a bona fide RSS and a cryptic RSS or the insertion of signal circles into chromosomal loci via a V(D)J recombination-like mechanism. Although wild-type, p53, p53 x scid, H2Ax, and ATM mutant thymocytes all showed similar levels of RSS end insertions

Topics: Abbreviations used, ds, double
Year: 2014
OAI identifier: oai:CiteSeerX.psu:
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