BLISS is a versatile and quantitative method for genome-wide profiling of DNA double-strand breaks
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Author(s)
Mirzazadeh, Reza
Garnerone, Silvano
Schneider, Martin W.
Kallas, Tomasz
Custodio, Joaquin
Wernersson, Erik
Bienko, Magda
Crosetto, Nicola
Yan, Winston Xia
Scott, David Arthur
Date Issued
May 2017
Journal
Nature Communications
Publisher
Nature Publishing Group
Citation
Yan, Winston X. et al. “BLISS Is a Versatile and Quantitative Method for Genome-Wide Profiling of DNA Double-Strand Breaks.” Nature Communications 8 (May 2017): 15058 © 2017 Macmillan Publishers Limited, part of Springer Nature
Version
Final published version
Abstract
Precisely measuring the location and frequency of DNA double-strand breaks (DSBs) along the genome is instrumental to understanding genomic fragility, but current methods are limited in versatility, sensitivity or practicality. Here we present Breaks Labeling In Situ and Sequencing (BLISS), featuring the following: (1) direct labelling of DSBs in fixed cells or tissue sections on a solid surface; (2) low-input requirement by linear amplification of tagged DSBs by in vitro transcription; (3) quantification of DSBs through unique molecular identifiers; and (4) easy scalability and multiplexing. We apply BLISS to profile endogenous and exogenous DSBs in low-input samples of cancer cells, embryonic stem cells and liver tissue. We demonstrate the sensitivity of BLISS by assessing the genome-wide off-target activity of two CRISPR-associated RNA-guided endonucleases, Cas9 and Cpf1, observing that Cpf1 has higher specificity than Cas9. Our results establish BLISS as a versatile, sensitive and efficient method for genome-wide DSB mapping in many applications.
MIT Department
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
McGovern Institute for Brain Research at MIT
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DOI of Published Version
https://doi.org/10.1038/ncomms15058
