BLISS is a versatile and quantitative method for genome-wide profiling of DNA double-strand breaks

Area of Science:

• Genomics
• Molecular Biology
• Biotechnology

Background:

• Accurate mapping of DNA double-strand breaks (DSBs) is crucial for understanding genomic instability.
• Existing methods for DSB detection have limitations in versatility, sensitivity, and practicality.

Purpose of the Study:

• To introduce Breaks Labeling In Situ and Sequencing (BLISS), a novel method for genome-wide DSB profiling.
• To demonstrate the sensitivity and efficiency of BLISS for analyzing DSBs in various biological samples.

Main Methods:

• BLISS involves direct labeling of DSBs in fixed cells or tissues.
• It utilizes linear amplification of tagged DSBs via in vitro transcription for low-input samples.
• Quantification is achieved using unique molecular identifiers, enabling scalability and multiplexing.

Main Results:

• BLISS was successfully applied to profile endogenous and exogenous DSBs in cancer cells, stem cells, and liver tissue.
• The method demonstrated high sensitivity in assessing genome-wide off-target activity of CRISPR-Cas9 and CRISPR-Cpf1 endonucleases.
• CRISPR-Cpf1 was observed to have higher specificity compared to CRISPR-Cas9.

Conclusions:

• BLISS is a versatile, sensitive, and efficient method for genome-wide DSB mapping.
• This technique offers significant advantages over existing methods for various research applications.
• BLISS facilitates detailed analysis of genomic fragility and DNA repair mechanisms.