Abstract
Backgrounds
The differences of DNA mutation patterns of genome editing by CRISPR (Clustered regularly interspaced Short Palindromic Repeats)-Cas9 and CRISPR-Cpf1 is not completely understood.
Methods
CRISPR-Cas9 or CRISPR-Cpf1 is applied for genome editing at the exact same four genomic locations in human embryonic kidney 293T (HEK293T) cells. The mutation patterns of the target genomic loci were analyzed by targeted deep sequencing.
Results
The mutation patterns of CRISPR-Cas9 and CRISPR-Cpf1 showed prominent differences. CRISPR-Cpf1 mediated genome engineering almost always resulted in deletion, while CRISPR-Cas9 showed remarkably high rate of insertion mutation of 1 base pair. The deletion patterns of CRISPR-Cas9 and CRISPR-Cpf1 were also different. The deletion patterns of CRISPR-Cas9 were composed of diverse and evenly distributed deletion patterns, while the mutation patterns of CRISPR-Cpf1 were composed of fewer major patterns.
Conclusion
Genome editing by CRISPR-Cas9 and CRISPR-Cpf1 shows different characteristics.
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Acknowledgements
This work was supported by the National Research Foundation (NRF) of Korea funded by the Ministry of Science & ICT (NRF-2017R1D1A1B030 35094, NRF-2017R1E1A1A01074529, NRF-2018M3A9H 3021707 to J.K.H., NRF-2017R1D1A1B03035760, NRF-2019R1C1C1010602 to J.W.H., NRF-2018R1A5A2023 879 to G.S., and 2014M3C7A1062896 to W.C.H.).
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T.B., J.W.H., and J.K.H designed the experiments and analyzed the results. T.B. conducted the experiments, W.C.H., D.L, and G.S conducted the computational analyses of the data. All authors contributed to writing of the manuscript.
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Taegeun Bae, Woo Chang Hwang, Dohyeon Lee, Giltae Song, Junseok W Hur, and Junho K Hur declare that they have no conflict of interest.
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This article does not include any research on humans and animals, and has it been carried out in accordance with the institutional and national guidelines.
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Bae, T., Hwang, W.C., Lee, D. et al. Characterization of distinct mutation patterns by CRISPR-Cas9 and CRISPR-Cpf1 genome editing systems. Mol. Cell. Toxicol. 15, 383–389 (2019). https://doi.org/10.1007/s13273-019-0041-9
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DOI: https://doi.org/10.1007/s13273-019-0041-9