Related Experiment Video
Updated: Jun 25, 2026
08:25
Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format
Published on: April 8, 2017
13.7K
High-Throughput Screening to Identify Novel Compounds Affecting the Genome Editing Efficiency of CRISPR System
Jiasong Chang1,2, Xiulong Yang1,2, Tong Zhang3
1Key Laboratory of Cellular Physiology, Shanxi Medical University, Ministry of Education, Taiyuan 030001, China.
Molecules (Basel, Switzerland)
|May 7, 2025
Summary
Researchers screened 9930 compounds to find those that modulate CRISPR/Cas9 gene editing. CP-724714 acts as a CRISPR decelerator, reducing off-target effects, while Clofarabine is a CRISPR accelerator, enhancing efficiency.
Area of Science:
- Biochemistry and Molecular Biology
- Pharmacology
- Genetic Engineering
Background:
- CRISPR/Cas9 gene editing offers therapeutic potential for genetic disorders but faces biosafety challenges due to off-target effects.
- Modulators of CRISPR/Cas9 efficiency, particularly those mitigating off-target mutations, are crucial for advancing gene therapy.
- Identifying such compounds can significantly improve the safety and efficacy of genome editing applications.
Purpose of the Study:
- To discover and characterize compounds that modulate the efficiency and specificity of the CRISPR/Cas9 system.
- To identify pharmacological tools that can reduce off-target effects in CRISPR/Cas9-mediated gene therapy.
- To explore compounds influencing DNA repair pathways, specifically single-strand annealing (SSA) repair.
Main Methods:
- Conducted a high-throughput screening of 9930 compounds in HEK 293FT cells to identify modulators of CRISPR/Cas9 activity.
- Performed two rounds of screening to validate compounds affecting genome editing efficiency.
- Investigated the impact of identified compounds on single-strand annealing (SSA) repair efficiency.
Main Results:
- Identified CP-724714, an ErbB2 (HER2) tyrosine kinase inhibitor, as a 'CRISPR decelerator' that decreases CRISPR/Cas9 efficiency and reduces off-target effects.
- Identified Clofarabine, a DNA synthesis inhibitor, as a 'CRISPR accelerator' that increases CRISPR/Cas9 efficiency.
- Discovered four compounds (Tranilast, Cerulenin, Rosolic acid, Resveratrol) affecting SSA repair, with Tranilast, Cerulenin, and Rosolic acid acting as decelerators, and Resveratrol as an accelerator.
Conclusions:
- CP-724714 and Clofarabine represent valuable tools for controlling CRISPR/Cas9 gene editing outcomes.
- Tranilast, Cerulenin, Rosolic acid, and Resveratrol offer new avenues for modulating SSA repair, a critical DNA repair pathway.
- These identified compounds hold significant promise for optimizing gene editing techniques and enhancing their therapeutic applicability in mammals.
