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Modulation of DNA double-strand break repair as a strategy to improve precise genome editing

Oncogene volume 39, pages 6393–6405 (2020)Cite this article

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Abstract

In the present day, it is possible to incorporate targeted mutations or replace a gene using genome editing techniques such as customisable CRISPR/Cas9 system. Although induction of DNA double-strand breaks (DSBs) by genome editing tools can be repaired by both non-homologous end joining (NHEJ) and homologous recombination (HR), the skewness of the former pathway in human and other mammals normally result in imprecise repair. Scientists working at the crossroads of DNA repair and genome editing have devised new strategies for using a specific pathway to their advantage. Refinement in the efficiency of precise gene editing was witnessed upon downregulation of NHEJ by knockdown or using small molecule inhibitors on one hand, and upregulation of HR proteins and addition of HR stimulators, other hand. The exploitation of cell cycle phase differences together with appropriate donor DNA length/sequence and small molecules has provided further improvement in precise genome editing. The present article reviews the mechanisms of improving the efficiency of precise genome editing in several model organisms and in clinics.

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Fig. 1: Cartoon depicting various steps associated with different DNA double-strand break repair pathways in mammals.
Fig. 2: Advancement of genome editing tools over the years and mechanism of genome editing using various tools.
Fig. 3: Cartoon showing a comparison of genome editing tools and their properties/advantages.
Fig. 4: Improvement in precise genome editing by utilising small molecules and overexpression of HR proteins with CRISPR/Cas9.
Fig. 5: Different forms and derivatives of SCR7.

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Acknowledgements

We thank Urbi Roy, Dipayan Ghosh and other members of SCR laboratory for critical reading and comments on the paper. This work was supported by grants from CEFIPRA (IFC/5203-4/2015/131), DAE (21/01/2016-BRNS/35074), DBT Glue-Grant (BT/PR23078/MED/29/1253/2017), IISc-DBT partnership programme (BT/PR27952-INF/22/212/2018) to SCR. UR is supported by Senior Research Fellowship (SRF) from CSIR, India.

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  1. Department of Biochemistry, Indian Institute of Science, Bangalore, 560012, India

    Ujjayinee Ray & Sathees C. Raghavan

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  1. Ujjayinee Ray
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Ray, U., Raghavan, S.C. Modulation of DNA double-strand break repair as a strategy to improve precise genome editing. Oncogene 39, 6393–6405 (2020). https://doi.org/10.1038/s41388-020-01445-2

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