期刊
NATURE COMMUNICATIONS
卷 8, 期 -, 页码 -出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/ncomms15058
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资金
- National Institute of General Medical Sciences [T32GM007753]
- Paul and Daisy Soros Fellowship
- Swedish Society for Medical Research (SSMF) Postdoctoral Fellowship
- NIH through NIMH [5DP1-MH100706, 1R01-MH110049]
- NSF
- New York Stem Cell Foundation
- Howard Hughes Medical Institute
- Simons Foundation
- Paul G. Allen Family Foundation
- Vallee Foundation
- Skoltech-MIT Next Generation Program
- Science for Life Laboratory
- Swedish Research Council [621-2014-5503, 521-2014-2866]
- Ragnar Soderberg Foundation
- Karolinska Institutet
- Karolinska Institutet Strategic Programme in Cancer
- Swedish Cancer Research Foundation [CAN 2015/585]
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.
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