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J Immunol Methods. 2012 Jan 31;375(1-2):176-81. doi: 10.1016/j.jim.2011.10.007. Epub 2011 Oct 18.

Translocation capture sequencing: a method for high throughput mapping of chromosomal rearrangements.

Author information

1
Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA.

Abstract

Chromosomal translocations require formation and joining of DNA double strand breaks (DSBs). These events disrupt the integrity of the genome and are involved in producing leukemias, lymphomas and sarcomas. Translocations are frequent, clonal and recurrent in mature B cell lymphomas, which bear a particularly high DNA damage burden by virtue of activation-induced cytidine deaminase (AID) expression. Despite the ubiquity of genomic rearrangements, the forces that underlie their genesis are not well understood. Here, we provide a detailed description of a new method for studying these events, translocation capture sequencing (TC-Seq). TC-Seq provides the means to document chromosomal rearrangements genome-wide in primary cells, and to discover recombination hotspots. Demonstrating its effectiveness, we successfully estimate the frequency of c-myc/IgH translocations in primary B cells, and identify hotspots of AID-mediated recombination. Furthermore, TC-Seq can be adapted to generate genome-wide rearrangement maps in any cell type and under any condition.

PMID:
22033343
PMCID:
PMC3285106
DOI:
10.1016/j.jim.2011.10.007
[Indexed for MEDLINE]
Free PMC Article

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