
Simultaneous Inhibition of Molecules Can Help Boost Precision and Efficiency of CRISPR
August 30, 2023 |
Researchers identified two molecule inhibitions that can enhance the precision and efficiency of gene editing. The results of their study can help advance the usage of CRISPR techniques in various applications.
CRISPR is a widely used tool for research all over the world. However, this technique has drawbacks, such as unintended mutations and low efficiency of selected DNA integration. To address this issue, the scientists analyzed a large-scale library of small molecules to choose targets that will be used to boost targeted genome insertions.
The results of the study showed that DNA-dependent protein kinase (DNA-PK) is the most effective target to enhance insertions in CRISPR-Cas9. The researchers extensively characterized AZD7648 and found out that the selective DNA-PK inhibitor can greatly boost the precision of gene editing. DNA polymerase theta (Polϴ) is another molecule that should be inhibited to improve the precision and efficiency of gene editing.
The researchers created a strategy called 2iHDR, which is the combination of the inhibition of DNA-PK and Polϴ. This technique enhanced templated insertions to 80% efficiency, as well as reduced unintended insertions and deletions to a minimum. The strategy also minimizes Cas9's off-target effects, which significantly improves the performance and fidelity of CRISPR-Cas9 gene editing.
For more information, read the journal article at Nature Communications.
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