BLESS

Breaks labeling and Enrichment on Streptavidin and Sequencing

BLESS is a genome-wide approach to map DSBs at nucleotide resolution (Crosetto et al., 2013). BLESS is able to detect telomere ends, Sce endonuclease_induced DSBs, and complex genome-wide DSB landscapes.In this method, DSBs are ligated in situ to a proximal linker covalently linked to biotin. The gDNA is extracted and fragmented, and the labeled fragments are captured on streptavidin beads. Next, a distal linker is ligated to the free end of the captured fragments. The fragments are released by linker digestion with I-SceI and PCR-amplified

Advantages:

  • Detects DSBs at nucleotide resolution
  • Does not depend on proteins that bind to DSBsãa source of bias
  • Does not depend on single-stranded DNA (ssDNA)ãa source of bias

Disadvantages:


Reagents:

Illumina Library prep and Array Kit Selector



Reviews:

Lee C. M., Cradick T. J., Fine E. J. and Bao G. Nuclease Target Site Selection for Maximizing On-target Activity and Minimizing Off-target Effects in Genome Editing. Mol Ther. 2016;24:475-487



References:

Ran F. A., Cong L., Yan W. X., et al. in vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015;520:186-191