High-Throughput Genome-Wide Translocation Sequencing
HTGTS was developed to study translocation mechanisms in mammalian cells (Chiarle et al., 2011). This approach is particularly suitable for studying for AID-dependent IgH class-switching (HTGTS-Rep-seq) (Lin et al., 2016) and CRISPR/Cas9 genome modifications (Mei et al., 2016).
In HTGTS-Rep-seq, genomic DNA from B-cell populations is sonicated and linearly amplified with a biotinylated primer that anneals downstream of a J segment. The biotin-labeled single-stranded DNA products are enriched with streptavidin beads, and the 3ê ends are ligated to a bridge adaptor containing a 6-nucleotide UMI.
- Higher efficiency compared to whole-genome sequencing
- Underestimates the frequency of DSBs (Hu et al., 2016)
- Limited by chromatin accessibility (Kim et al., 2015)
Illumina Library prep and Array Kit Selector
None available yet
Schwer B., Wei P. C., Chang A. N., et al. Transcription-associated processes cause DNA double-strand breaks and translocations in neural stem/progenitor cells. Proc Natl Acad Sci U S A. 2016;113:2258-2263
Frock R. L., Hu J., Meyers R. M., Ho Y. J., Kii E. and Alt F. W. Genome-wide detection of DNA double-stranded breaks induced by engineered nucleases. Nat Biotechnol. 2015;33:179-186