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Role of RS-1 derivatives in homology-directed repair at the human genome ATG5 locus

Archives of Pharmacal Research 2020년 43권 6호 p.639 ~ 645
전인숙 ( Jeon In-Sook ) - Chungbuk National University College of Medicine Department of Biochemistry

신재천 ( Shin Jae-Cheon ) - Pohang Center for Evaluation of Biomaterial
김승렬 ( Kim Seung-Ryul ) - Chungbuk National University College of Medicine Department of Biochemistry
박관식 ( Park Kwan-Sik ) - Chungbuk National University College of Medicine Department of Biochemistry
유현정 ( Yoo Hyun-Jung ) - Chungbuk National University Department of Consumer Science
이광열 ( Lee Kwang-Youl ) - Chonnam National University College of Pharmacy
 ( Lee Hak-Kyo ) - Chonbuk National University Department of Animal Biotechnology
최중국 ( Choi Joong-Kook ) - Chungbuk National University College of Medicine Department of Biochemistry

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.

키워드

ATG5; CRISPR/Cas9; Homology directed repair; RS-1
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