Related Experiment Video
Updated: May 17, 2026
Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells
Published on: May 30, 2025
Simultaneous CRISPR/Cas9-induced double-strand breaks are lethal in models of pancreatic cancer
Selina Shiqing K Teh1, Akhil Kotwal2, Alexis Bennett1
1Department of Pathology, The Sol Goldman Pancreatic Cancer Research Center.
Abstract:
While radiation is an effective oncologic therapy, killing cancer by inducing DNA double-strand breaks (DSBs), it lacks specificity for neoplastic cells. We have previously adapted the CRISPR/Cas9 gene-editing technology as a cancer-specific treatment modality targeting somatic mutations in pancreatic cancer (PC). However, its tumoricidal potential remains unclear, especially in comparison with therapeutic doses of radiation. Here, we demonstrate that CRISPR/Cas9-induced DSBs are more cytotoxic in PCs than a comparable number of radiation-induced DSBs. We observed more than 90% tumor growth inhibition by targeting 9 sites with cancer-specific sgRNAs. Through both bioinformatics and cytogenetics analyses, we found that CRISPR/Cas9-induced DSBs triggered ongoing chromosomal rearrangements, with 87% of structural variants not directly produced from the initial CRISPR/Cas9-induced DSBs, and chromosomal instability peaking before cell death. By comparing the cytotoxicity of CRISPR/Cas9- and radiation-induced DSBs, we demonstrated that the number of DSBs required to achieve equitoxic effects was approximately 3 times higher for radiation than CRISPR/Cas9. Finally, we showed that PC cells that had survived CRISPR/Cas9 targeting retained susceptibility to subsequent CRISPR/Cas9-induced DSBs at different genomic sites with more than 87% growth inhibition. Together, our data support the therapeutic potential of CRISPR/Cas9 as an anticancer strategy.
Insights
CRISPR/Cas9 gene editing induces more DNA double-strand breaks (DSBs) in pancreatic cancer cells than radiation. This cancer therapy shows over 90% tumor growth inhibition and potential for repeated treatments.
Area of Science:
- Molecular biology
- Genetics
- Cancer research
Background:
- Radiation therapy is an effective cancer treatment but lacks specificity.
- CRISPR/Cas9 gene editing has been adapted for cancer therapy.
- The tumoricidal potential of CRISPR/Cas9 compared to radiation is unclear.
Purpose of the Study:
- To compare the cytotoxicity of CRISPR/Cas9-induced DNA double-strand breaks (DSBs) with radiation-induced DSBs in pancreatic cancer.
- To investigate the genomic consequences of CRISPR/Cas9-induced DSBs.
- To assess the efficacy of repeated CRISPR/Cas9 treatments.
Main Methods:
- Utilized CRISPR/Cas9 gene editing targeting somatic mutations in pancreatic cancer.
- Quantified DNA double-strand breaks (DSBs) induced by CRISPR/Cas9 and radiation.
- Performed bioinformatics and cytogenetics analyses to assess chromosomal rearrangements.
- Compared tumor growth inhibition and cell death rates between CRISPR/Cas9 and radiation treatments.
- Evaluated the efficacy of sequential CRISPR/Cas9 treatments on surviving cells.
Main Results:
- CRISPR/Cas9-induced DSBs were more cytotoxic than radiation-induced DSBs.
- Targeting 9 sites with cancer-specific sgRNAs resulted in over 90% tumor growth inhibition.
- CRISPR/Cas9-induced DSBs triggered chromosomal rearrangements, with 87% not directly from initial breaks.
- Chromosomal instability peaked before cell death.
- Approximately 3 times more DSBs were required for radiation than CRISPR/Cas9 to achieve equitoxic effects.
- Surviving cells retained susceptibility to subsequent CRISPR/Cas9 treatments, showing >87% growth inhibition.
Conclusions:
- CRISPR/Cas9 gene editing demonstrates superior cytotoxicity compared to radiation for pancreatic cancer.
- CRISPR/Cas9 induces complex genomic instability, contributing to its tumoricidal effect.
- CRISPR/Cas9 represents a promising, potentially repeatable, anticancer therapeutic strategy.
