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Updated: Jun 6, 2026

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
Published on: March 31, 2022
Pan-Cancer Genomic Scars of Alternative End Joining and Single-Strand Annealing
Ashini Modi1, Alessandro Zito2,3, Giovanni Parmigiani2,3
1Harvard College, Harvard University, Cambridge, MA, USA.
DNA double-strand breaks (DSBs) are repaired by homologous recombination (HR) or backup pathways like single-strand annealing (SSA) and alternative end joining (Alt-EJ). This study reveals these backup pathways are influenced by genomic context, not just HR deficiency.
Area of Science:
- Genomics
- Molecular Biology
- Cancer Research
Background:
- DNA double-strand breaks (DSBs) are critical DNA lesions that can lead to genome instability and cancer.
- Homologous recombination (HR) is the primary high-fidelity repair pathway for DSBs.
- Error-prone pathways, single-strand annealing (SSA) and alternative end joining (Alt-EJ), act as backups, particularly in HR-deficient cells.
Purpose of the Study:
- To systematically analyze the genomic scars left by SSA and Alt-EJ across various cancer types.
- To investigate the relationship between these backup repair pathways and HR deficiency.
- To understand the influence of local genomic and transcriptional context on DSB repair pathway engagement.
Main Methods:
- Whole-genome sequencing of 2,157 tumors across 17 cancer types.
- Identification and characterization of SSA-like and Alt-EJ-like deletion signatures.
- Correlation analysis between deletion burdens, HR deficiency scores, and genomic features.
Main Results:
- Identified 832 SSA-like and 37,359 Alt-EJ-like deletions.
- Alt-EJ is the dominant backup pathway in HR-deficient tumors.
- Elevated SSA-like deletions were observed in prostate adenocarcinoma and hepatocellular carcinoma, irrespective of HR deficiency.
- SSA-like deletions show enrichment in SINE-rich regions and near transcription start sites in specific tumor types.
Conclusions:
- SSA and Alt-EJ repair pathways are active beyond HR-deficient contexts.
- The engagement of these backup pathways is significantly influenced by local genomic and transcriptional environments.
- Genomic scars provide insights into DSB repair mechanisms beyond HR deficiency, revealing distinct repair activities.
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Fixing Double-strand Breaks
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