Breaks Labeling in situ and sequencing (BLISS)

• DOI: 10.21203/rs.2.1448/v2
• Publication Date: 2019-11-13T11:51:22Z
• Authors: Winston X. Yan; Reza Mirzazadeh; Silvano Garnerone; David Scott; Martin W. Schneider; Tomasz Kallas; Joaquin Custodio; Yinqing Li; Linyi Gao; Yana Federova; Erik Wernersson; Bernd Zetsche; Feng Zhang; Magda Bienko; Nicola Crosetto
• Keywords: BLISS

Abstract Precisely measuring the location and frequency of DNA double-strand breaks \(DSBs) along genome is instrumental to understanding genomic fragility, but current methods are limited in versatility, sensitivity, or practicality. Here, we present Breaks Labeling _In Situ_ Sequencing \(BLISS), featuring: 1) direct labeling DSBs fixed cells tissue sections on a solid surface; 2) low-input requirement by linear amplification tagged _in vitro_ transcription; 3) quantification through unique molecular identifiers; 4) easy scalability multiplexing. We apply BLISS profile endogenous exogenous samples cancer cells, embryonic stem liver tissue. demonstrate sensitivity assessing genome-wide off-target activity two CRISPR-associated RNA-guided endonucleases, Cas9 Cpf1, observing that Cpf1 has higher specificity than Cas9. Our results establish as versatile, sensitive, efficient method for DSB mapping many applications. W.Yan R.Mirzazadeh equally contributed this work.