Abstract
Genome editing is a useful tool in basic and clinical research. Among the several approaches used in genome editing, the CRISPR–Cas9 system using clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) along with a guide RNA has been developed recently. The CRISPR/Cas9 system induces site-specific double-stranded DNA breaks, which result in DNA repair via non-homologous end joining (NHEJ) or homology-directed repair (HDR). However, HDR efficiency is lower than that of NHEJ and accordingly poses a challenge in genome modification studies. Several chemical compounds including RS-1 have been shown to enhance the HDR knock-in process by two- to six-fold in HEK 293 cells and rabbit embryos. Based on this finding, we developed an antibiotic resistance system to screen RS-1 chemical derivatives, which may promote efficient HDR. In this study, we report several chemical compounds with high knock-in efficiency at the ATG5 gene locus, using HeLa cell-based assays.
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Acknowledgements
This work was supported by (1) Next Generation BioGreen21 project (Project Nos. PJ01334802 & PJ01315101) of the Rural Development Administration, Korea, (2) Ministry of Education and the National Research Foundation of Korea (NRF-2017S1A5B8059946), and (3) Korea Ministry of Environment (MOE) and Korea Environment Industry & Technology Institute (KEITI) as a "Technology Program for establishing biocide safety management” [2018002490003]. The chemical library used in this study was kindly provided by Korea Chemical Bank (http://www.chembank.org/) of KRICT.
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Jeon, IS., Shin, JC., Kim, S.R. et al. Role of RS-1 derivatives in homology-directed repair at the human genome ATG5 locus. Arch. Pharm. Res. 43, 639–645 (2020). https://doi.org/10.1007/s12272-020-01226-1
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DOI: https://doi.org/10.1007/s12272-020-01226-1